The DOI matches its Crossref record; 61 of the 61 references it cites were found in a registry or an index; 11 of the 29 sources its claims cite were checked for support.
Improving mRNA-Based Therapeutic Gene Delivery by Expression-Augmenting 3′ UTRs Identified by Cellular Library Screening
Open accessAbstract
Synthetic mRNA has emerged as a powerful tool for the transfer of genetic information, and it is being explored for a variety of therapeutic applications. Many of these applications require prolonged intracellular persistence of mRNA to improve bioavailability of the encoded protein. mRNA molecules are intrinsically unstable and their intracellular kinetics depend on the UTRs embracing the coding sequence, in particular the 3' UTR elements. We describe here a novel and generally applicable cell-based selection process for the identification of 3' UTRs that augment the expression of proteins encoded by synthetic mRNA. Moreover, we show, for two applications of mRNA therapeutics, namely, (1) the delivery of vaccine antigens in order to mount T cell immune responses and (2) the introduction of reprogramming factors into differentiated cells in order to induce pluripotency, that mRNAs tagged with the 3' UTR elements discovered in this study outperform those with commonly used 3' UTRs. This approach further leverages the utility of mRNA as a gene therapy drug format.
Overview
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Written from the paper’s full text.
TL;DR
The authors adapted SELEX into a cell-culture-based selection process in human dendritic cells to discover naturally occurring 3′ UTR elements that stabilize synthetic mRNA, identifying mtRNR1 and AES motives as top performers. mRNAs tagged with AES-mtRNR1 or mtRNR1-AES 3′ UTRs outperformed the standard double β-globin (2hBg) UTR in antigen-encoding mRNA vaccination in mice and in mRNA-based reprogramming of human fibroblasts to iPSCs.
Key findings
- Six selection rounds in Actinomycin D–treated immature hDCs enriched stabilizing sequences, with the round-five library showing an approximately 6-fold increase in half-life versus the starting library; >50% (n = 216) of 402 sequenced clones derived from only 15 endogenous transcripts, including FCGRT, LSP1, CCL22, HLA-DRB4, CCL3, AES, PLD3, and mtRNR1.
- All tested 3′ UTRs outperformed the commonly used human β-globin (hBg) UTR; dUTRs containing mtRNR1, particularly with AES or hBg, were moderately superior to 2hBg. Functional half-lives correlated negatively with predicted miRNA-binding site number and positively with total hybridization energy.
- In murine C2C12 myoblasts all tested 3′ UTRs outperformed hBg by 2.3- to 3.6-fold; in human foreskin fibroblasts mtRNR1/AES combinations were superior to hBg by 2.4- to 3.5-fold.
- In BALB/c mice (n = 8) given i.v. liposome-formulated luciferase mRNA, AES-mtRNR1 tagging gave higher and more prolonged splenic expression than 2hBg; mice (n = 5) immunized three times with gp70 mRNA carrying AES-mtRNR1 generated significantly higher frequencies of gp70-specific CD8+ T cells. In reprogramming, AES-mtRNR1 yielded up to 60-fold higher colony-forming units versus 6.5-fold for 2hBg, and two transfection rounds sufficed for iPSC colony formation.
Methods
- Cell-based selection: mRNA purified from ActD-treated immature hDCs was fragmented to 100–750 nt, reverse transcribed, and cloned as 3′ UTRs between destabilized EGFP (d2eGFP) and a 60-adenosine poly(A) tail; six rounds of electroporation into hDCs, incubation, recovery, and amplification, with decay curves by qRT-PCR.
- Functional characterization: 15 core motives cloned as 3′ UTRs upstream of Luc2CP (PEST-domain luciferase) or Luc reporters, transfected into hDCs, C2C12, and HFFs; protein expression kinetics, functional half-life, and area under the protein decay curve measured; miRNA-binding sites predicted with IntaRNA.
- Applications: in vivo vaccination in BALB/c mice with liposome-formulated mRNA (10 μg Luc mRNA for imaging; 5 μg gp70/AH1-A5 mRNA on days 0, 7, 14 for tetramer-based CD8+ T cell quantification); iPSC reprogramming of HFFs by daily lipofection of OSKMNL plus EKB mRNAs with miRNAs 302a-d/367, assessed by alkaline phosphatase staining, TRA-1-81, and qRT-PCR of endogenous markers.
Limitations
- Stated: it was not the scope of the study to test the discovered elements across a broader range of cell types, and because mRNA-stability regulators are cell type specific, there may be cell types in which AES-mtRNR1-based elements are not superior to conventional 3′ UTRs; future studies must explore whether the elements improve expression and immune responses for mRNA delivered by other routes.
- Stated: homologous dUTRs were not analyzed because they are prone to recombination, which is noted as a drawback of the 2hBg 3′ UTR.
- Evident from the text: animal experiments used small groups (n = 8 for imaging, n = 5 for immunogenicity) with no statistical pre-determination of sample size, and several authors are employees of or consultants for BioNTech RNA Pharmaceuticals and inventors on related patent applications.
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Claims the paper makes
29 Own result · 10 Not checked yet · 10 Own interpretation · 7 Own method · 3 Supported · 3 Partly supported · 3 No basis given · 2 No full text of the sources · 1 Supported so far · 1 Couldn’t confirm · accuracy not yet measured
Every source the paper cites for this claim was checked and supports it.
In the paper
“We had previously shown in hDCs that two head-to-tail-cloned hBg 3′ UTRs (2hBg) are superior to a single copy in improving mRNA stability.7”
What the sources say
- Supportedcites [7] Holtkamp, 2006, Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells, Blood source
The section on tandem human β-globin 3′ UTRs reports that two head-to-tail copies outperform one and identifies message stability as the primary effect in immature dendritic cells. The Discussion also states that two copies are better than one at improving RNA translational characteristics. (LLM, conf=0.90)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
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- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“mRNAs with longer 3′ UTRs have a shorter half-life22”
What the sources say
- Supportedcites [22] Schwanhäusser, 2011, Global quantification of mammalian gene expression control, Nature source
In “Estimating the impact of post-transcriptional, translational and post-translational control on protein abundance” (p. 10), the paper reports that mRNAs with long 3′ UTRs are, on average, less stable. This supports the claimed relationship between 3′ UTR length and mRNA half-life as an average trend, not a rule for every transcript. (LLM, conf=0.94)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“This is in line with reports that mRNA translation decreases with a higher number of miRNAs binding to the 3′ UTR.45”
What the sources say
- Supportedcites [45] Jackson, 2010, The mechanism of eukaryotic translation initiation and principles of its regulation, Nat. Rev. Mol. Cell Biol. source
The “Translation regulation by miRNAs” section says repression increases as more miRNAs associate with the 3′ UTR. It also identifies repression of mRNA translation as one component of that effect. (LLM, conf=0.91)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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- Supported
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“We describe here a novel and generally applicable cell-based selection process for the identification of 3′ UTRs that augment the expression of proteins encoded by synthetic mRNA.”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Within each alignment, we identified a core area present in all clones (defined as motives) and flanked by up- and downstream areas of different length (Tables S2 and S3).”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Functional half-lives of mRNAs correlated negatively with the number of predicted miRNA-binding sites in the 3′ UTRs with which they were equipped (Figure 2C)”
plaincorrelational- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Sequence alignment analysis of these individual clones revealed that the majority of them mapped, at least partially, to the 5′ or 3′ UTR of coding mRNAs (Table 1).”
plainn = 402- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Almost all dUTRs outperformed hBg (Figure 2B; >1.00)”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“All tested 3′ UTRs were found to perform better than the commonly used standard hBg.”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“All transcripts except for one are abundantly expressed in hDCs (Table 1; Table S2).”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Comparing the half-life of the round five library to the starting mRNA library, we found an about 6-fold increase (Table S1).”
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“whereas they correlated positively with the total hybridization energies as a surrogate for impaired miRNA binding (Figure 2D).”
plaincorrelational- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Compared to the percentage of reads that could be aligned to these 15 core areas from NGS data of the starting library, all motives were highly enriched during the selection process (Table 1).”
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“dUTRs carrying the mtRNR1 element, in particular in combination with AES and hBg motives, were moderately superior to 2hBg (Figure 2B).”
plaincorrelational- Asserted: passes — The authors put it forward as true.
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In the paper
“Six of the motives mapped to mRNAs (AES, PLD3, PTRF, CCDC124, PTMA, and MYH9), which are known to have a high median mRNA half-life.22”
What the sources say
- Not checked yetcites [22] Schwanhäusser, 2011, Global quantification of mammalian gene expression control, Nature source
boostedcorrelational- Asserted: passes — The authors put it forward as true.
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- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In the murine myoblast cell line C2C12, all tested 3′ UTRs significantly outperformed hBg by 2.3- to 3.6-fold (Figure 3C).”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Over 50% of the sequences enriched by several selection rounds were represented by 15 distinct motives defined by identical core areas but diverse 5′/3′ ends.”
plain- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“More importantly, even when cells were transfected only two times within 48 hr with mRNAs containing the UTRs discovered in this study, colony formation of iPSCs was robustly detectable from day 8 on.”
- Also stated in: § Discussion · plain
“With mRNAs tagged with the AES-mtRNR1 3′ UTR, two transfection rounds were sufficient for the generation of iPSC colonies from human neonatal fibroblasts.”
plaindirect- Asserted: passes — The authors put it forward as true.
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- Also stated in: § Discussion · plain
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“we developed a cell culture-based method for the systematic enrichment of naturally occurring RNA sequences that improve the kinetics of gene-encoding mRNA in vitro and in vivo when used as 3′ UTRs.”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“which are insufficient for 2hBg-tagged reprogramming factors (Figure 5A).”
plain- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Mice immunized with AES-mtRNR1 3′ UTR-tagged gp70 generated a significantly higher frequency of gp70 antigen-specific CD8+ T cells in the spleen as compared to mice immunized with the 2hBg-tagged mRNA (Figure 4B).”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Here the mtRNR1-AES 3′ UTR performed best by achieving significantly higher maximum protein levels (Figures S3C and S3D).”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Of note, the activity of the pluripotency-associated TRA-1-81 and AP markers in mtRNR1-AES- or AES-mtRNR1-derived iPSC colonies and on the cell level was comparable to that of 2hBg-derived colonies.”
plain- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The AES-mtRNR1 and mtRNR1-AES combination motives that we determined functionally as the best performers have a far lower number of putative miRNA-binding sites as compared to the other dUTRs we had discovered and tested.”
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In human foreskin fibroblasts (HFFs), mtRnR1- and AES-element combinations were superior to hBg by 2.4- to 3.5-fold (Figure 3D).”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
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A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The 3′ UTRs discovered by this approach improve therapeutic mRNA-based gene transfer for various applications.”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
In the paper
“whereas mRNAs with shorter 3′ UTRs are less efficiently translated.23”
What the sources say
- Not checked yetcites [23] Tanguay, 1996, Translational efficiency is regulated by the length of the 3′ untranslated region, Mol. Cell. Biol. source
plaindirect- Asserted: passes — The authors put it forward as true.
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A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Optimized 3′ UTRs further increase the efficiency of a potent, simple, safe, and good manufacturing practice (GMP)-compatible gene delivery by mRNA for integration-free iPSC generation from easily accessible adult human somatic cells.”
plaindirect- Asserted: passes — The authors put it forward as true.
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A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The optimized expression translated into stronger induction of antigen-specific immune responses in the vaccinated mice.”
plaindirect- Asserted: passes — The authors put it forward as true.
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In the paper
“One of the implications of this improvement is to make refractory cell lines or cell types like EPCs that require even more rounds of transfections amenable to iPSC generation.”
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
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A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“As a consequence, with conventional synthetic libraries, relevant sequences may be lost during electroporation.”
hedgedconditional- Asserted: passes — The authors put it forward as true.
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- Probably not a substantial claim. 23
- It may state two separate claims as one. 22
- The quote only partly states this claim. 1
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“The potency of the antigen-specific immune response correlates positively with the persistence of the synthetic mRNA in the hDCs and the amount of translated protein.7,8”
What the sources say
- Supportedcites [7] Holtkamp, 2006, Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells, Blood source
The Discussion links RNA stability and translation to sustained antigen presentation and improved T-cell stimulation. The results on HCMV pp65 further report stronger expansion of human antigen-specific T cells with the stabilized RNA construct. These passages support the claimed positive relationship, though the paper describes it qualitatively rather than as a calculated correlation. (LLM, conf=0.87)
- Could not verifycites [8] Princiotta, 2003, Quantitating protein synthesis, degradation, and endogenous antigen processing, Immunity sourceExperimental — accuracy not yet measured
The Abstract and the passage headed “Quantitating Antigen Processing in Professional Antigen-Presenting Cells” quantify viral-protein synthesis and MHC class I peptide-complex formation in mouse cells. They do not address synthetic mRNA persistence in human dendritic cells or test whether either mRNA persistence or translated-protein amount correlates with the potency of an antigen-specific immune response (searched: synthetic mRNA persistence, mRNA stability, and T-cell activation). (LLM, conf=0.76)
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Robust induction of an immune response by a cancer vaccine requires efficient expression of the vaccine antigen-encoding mRNA specifically in DCs to achieve a high density of epitope-loaded MHCs.1,42”
What the sources say
- Partly supportedcites [1] Sahin, 2014, mRNA-based therapeutics--developing a new class of drugs, Nat. Rev. Drug Discov. sourceExperimental — accuracy not yet measured
The Cancer immunotherapy section reports that mRNA was selectively taken up by lymph-node dendritic cells, that its antigen was presented on MHC molecules, and that this approach induced strong antigen-specific T-cell responses in animal models. It does not establish that robust responses require expression specifically in dendritic cells or a high density of epitope-loaded MHC molecules. (LLM, conf=0.86)
- Supportedcites [42] Kuhn, 2011, Determinants of intracellular RNA pharmacokinetics: Implications for RNA-based immunotherapeutics, RNA Biol. source
The “Determinants of intracellular RNA pharmacokinetics” section (page unavailable) describes antigen translation in dendritic cells as the mode of action of RNA vaccines and reports a direct relationship between antigen produced in those cells and the density of surface peptide–MHC molecules for T-cell stimulation. The “Characteristics of RNA to be Used for Cancer Immunotherapy” section calls efficient translation of antigen-encoding RNA pivotal to generating effective immune responses. (LLM, conf=0.90)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Protein expression kinetics, which integrate all functionally relevant translational characteristics of mRNA, were measured.23,42”
What the sources say
- Partly supportedcites [23] Tanguay, 1996, Translational efficiency is regulated by the length of the 3′ untranslated region, Mol. Cell. Biol. sourceExperimental — accuracy not yet measured
The Results (Fig. 1 and Table 2) show that protein-expression kinetics capture translational efficiency and functional mRNA stability, which together affect protein yield. The paper does not establish that those kinetics integrate *all* functionally relevant translational characteristics of mRNA; its analysis concerns the measured luciferase constructs in CHO cells. (LLM, conf=0.82)
- Supportedcites [42] Kuhn, 2011, Determinants of intracellular RNA pharmacokinetics: Implications for RNA-based immunotherapeutics, RNA Biol. source
In “Determinants of intracellular RNA pharmacokinetics,” the paper describes measuring reporter-protein expression over time as an integrated record of relevant mRNA characteristics. It says the resulting curves allow translational efficiency, functional mRNA stability, and total protein expression to be deduced. (LLM, conf=0.92)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
- Partly supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“A general challenge of cell culture-based enrichment and selection methods is the delivery of the mRNA pool.43”
What the sources say
- Could not verifycites [43] Thiel, 2011, Nucleotide bias observed with a short SELEX RNA aptamer library, Nucleic Acid Ther. sourceExperimental — accuracy not yet measured
The Results and Targeted and nontargeted SELEX sections describe selecting RNA aptamers against cultured cells, but do not discuss delivering an mRNA pool. Searches for “delivery of the mRNA pool” and for cell-culture delivery or uptake challenges found no passage addressing the claimed general challenge. Page numbers were not available in our copy. (LLM, conf=0.77)
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Could not verify
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“Second, we studied the efficiency and robustness of cellular reprogramming of human fibroblasts to iPSCs with mRNA-encoded transcription factors, which is considered to benefit from prolonged mRNA stability.17,51”
What the sources say
- Not checked — no full text of the sourcecites [17] Poleganov, 2015, Efficient Reprogramming of Human Fibroblasts and Blood-Derived Endothelial Progenitor Cells Using Nonmodified RNA for Reprogramming and Immune Evasion, Hum. Gene Ther. source
- Supportedcites [51] Warren, 2010, Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA, Cell Stem Cell source
In Results, under “Development of modified-RNAs for directing cell fate” (page not available), the authors say they designed the mRNA to boost its cytoplasmic half-life. They also explain that sustained expression of reprogramming factors is needed over days to weeks, supporting the stated benefit of prolonged mRNA stability. Their protocol additionally used repeated transfection to sustain expression. (LLM, conf=0.88)
plainconditional- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Of note, the mtRNR1 and AES motives were among the 3′ UTRs that had the lowest number of predicted binding sites for miRNAs and the highest total hybridization energies.”
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The average stability of the RNA pool increased in each round and reached a plateau after round four, indicating an enrichment of stabilizing 3′ UTR sequences.”
hedgeddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Of note, as many 5′ as 3′ UTRs were found in the most stable transcripts.”
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Moreover, we show, for two applications of mRNA therapeutics, namely, (1) the delivery of vaccine antigens in order to mount T cell immune responses and (2) the introduction of reprogramming factors into differentiated cells in order to induce pluripotency, that mRNAs tagged with the 3′ UTR elements discovered in this study outperform those with commonly used 3′ UTRs.”
boosteddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
In the paper
“Moreover, in line with published data, the length of the 3′ UTR elements correlated negatively with their stabilizing effect (Figure S2).22”
What the sources say
- Not checked yetcites [22] Schwanhäusser, 2011, Global quantification of mammalian gene expression control, Nature source
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“As a consequence, the total amount of protein produced over time (measured as area under the protein decay curve) in hDCs by mRNAs with all tested 3′ UTRs was significantly higher in comparison to hBg and particularly pronounced for the mtRNR1-AES 3′ UTR (Figure 3B).”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“mRNAs containing the 3′ UTRs discovered in this study performed better than 2hBg 3′ UTR-tagged mRNAs, with the AES-mtRNR1 3′ UTR variant being significantly superior (up to 60-fold higher colony-forming units as compared to 6.5-fold), as shown by alkaline phosphatase (AP)-staining of colonies on day 9 (Figures 5B and 5C).”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The selection rounds enriched for motives mapping to the UTR of the respective mRNAs, but not to their on average much longer coding regions.”
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Functional mRNA half-life and time to maximum protein expression, which are both impacted by mRNA stability, were significantly enhanced across all tested 3′ UTRs (Figures S3A and S3B).”
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one. The quote only partly states this claim.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The hBg 3′ UTR mediated the lowest protein expression, whereas the dUTRs combining mtRNR1 and AES motives’ protein expression was augmented and prolonged (Figure 3A).”
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The colonies had distinct well-defined borders, consisted of tightly packed small cells with hESC morphology (Figure S4A), were highly active for AP (Figure S4B), and stained positive for the hESC surface marker TRA-1-81 (Figure S4B).”
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Splenic luciferase expression in mice injected with mtRNR1-AES or AES-mtRNR1 3′ UTR-tagged mRNA was higher and more prolonged as compared to mice injected with mRNA containing the 2hBg 3′ UTR. This difference was pronounced and significant within the first 24 hr, with AES-mtRNR1-tagged reporter mRNA being particularly superior (Figure 4A).”
- Also stated in: § Discussion · boosted
“By doing so, we demonstrated that AES-mtRNR1 and mtRNR1-AES tagging improved expression of the mRNA in mouse lymphatic compartments as compared to the 2hBg 3′ UTR.”
boosteddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Also stated in: § Discussion · boosted
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“This approach further leverages the utility of mRNA as a gene therapy drug format.”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may be too vague to check against evidence. The paper’s conclusions may not depend on it.
- Rests on an assumption: hDCs survive in cell culture for an extended period of time without cell division.
- Rests on an assumption: hDCs are the host cells for vaccine antigen delivery.
None of the sources cited for this claim have been checked yet.
In the paper
“Homologous dUTRs (fusion of two copies of the same element) were not analyzed, as they are prone to recombination and lead to undesired by-products,41”
What the sources say
- Not checked yetcites [41] Bradley, 1997, Recombinant DNA sequences generated by PCR amplification, Mol. Biol. Evol. source
Checked before — its earlier verdict is held back until a new check runs.
plaindirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
- Not checked yet
None of the sources cited for this claim have been checked yet.
In the paper
“The commonly used 3′ UTRs in mRNA therapeutics are derived from α- and β-globins,24,25”
What the sources say
- Not checked yetcites [24] Peixeiro, 2011, Control of human beta-globin mRNA stability and its impact on beta-thalassemia phenotype, Haematologica source
Checked before — its earlier verdict is held back until a new check runs.
- Not checked — no full text of the sourcecites [25] Waggoner, 2003, Regulation of alpha-globin mRNA stability, Exp. Biol. Med. (Maywood) source
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Not checked yet
In the paper
“These results are consistent with the mechanism of miRNA action, which is to inhibit the translation of their target mRNA and to mediate its degradation.40”
What the sources say
- Not checked — no full text of the sourcecites [40] Brümmer, 2014, MicroRNA binding sites in the coding region of mRNAs: extending the repertoire of post-transcriptional gene regulation, BioEssays source
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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Probably not a substantial claim. It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“globin 3′ UTRs are now being broadly used for mRNA delivery into various cell types.”
What the sources say
- Not checked — no full text of the sourcecites [27] Aviv, 1976, Biosynthesis and stability of globin mRNA in cultured erythroleukemic Friend cells, Cell source
- Not checked yetcites [28] Ross, 1985, Half-lives of beta and gamma globin messenger RNAs and of protein synthetic capacity in cultured human reticulocytes, Blood source
Checked before — its earlier verdict is held back until a new check runs.
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“mRNA-encoded vaccine antigens equipped with 2hBg generate superior immune responses and survival benefit in mouse tumor models,35–37”
What the sources say
- Not checked — no full text of the sourcecites [35] Diken, 2013, mTOR inhibition improves antitumor effects of vaccination with antigen-encoding RNA, Cancer Immunol. Res. source
- Not checked — no full text of the sourcecites [36] Kreiter, 2011, FLT3 ligand enhances the cancer therapeutic potency of naked RNA vaccines, Cancer Res. source
- Not checked yetcites [37] Van Nuffel, 2012, Dendritic cells loaded with mRNA encoding full-length tumor antigens prime CD4+ and CD8+ T cells in melanoma patients, Mol. Ther. source
Checked before — its earlier verdict is held back until a new check runs.
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
In the paper
“Protein-encoding mRNA has emerged as an attractive new technology for gene delivery.1”
What the sources say
- Not checked yetcites [1] Sahin, 2014, mRNA-based therapeutics--developing a new class of drugs, Nat. Rev. Drug Discov. source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Quality of purified mRNA was assessed by spectrophotometry (NanoDrop 2000c; VWR International, Darmstadt, Germany) and on-chip electrophoresis on the 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA).”
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“After 6 hr, only five mice could be followed up for the following time points, and one mouse was lost to follow-up after 24 hr.”
plainn = 5- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Sequence variability of the starting library was determined by NGS.”
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“Typically, only a small amount of mRNA pool resembling just a tiny fraction of the theoretical complexity of a randomized library can be delivered into the selection environment, which limits the discovery space substantially.44”
What the sources say
- Not checked yetcites [44] Javorovic, 2005, RNA transfer by electroporation into mature dendritic cells leading to reactivation of effector-memory cytotoxic T lymphocytes: a quantitative analysis, Mol. Ther. source
Checked before — its earlier verdict is held back until a new check runs.
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Not checked yet
None of the sources cited for this claim have been checked yet.
In the paper
“Alphaviral self-replicating RNA is an alternative vector for fast reprogramming, which requires only one transfection.52,53”
What the sources say
- Not checked yetcites [52] Yoshioka, 2017, Enhanced generation of iPSCs from older adult human cells by a synthetic five-factor self-replicative RNA, PLoS ONE source
- Not checked yetcites [53] Yoshioka, 2013, Efficient generation of human iPSCs by a synthetic self-replicative RNA, Cell Stem Cell source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
In the paper
“This formulation delivers mRNA systemically into lymphoid compartments, and it is selectively taken up by antigen-presenting cells.6”
What the sources say
- Not checked — no full text of the sourcecites [6] Kranz, 2016, Systemic RNA delivery to dendritic cells exploits antiviral defence for cancer immunotherapy, Nature source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Mouse studies were approved by the German regulatory authorities for animal welfare, and mice were kept in accordance with federal and state policies on animal research at BioNTech.”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“We mitigated this risk by constructing a library of reduced complexity from an amplified starting pool, thus ensuring representation of the full repertoire of sequences in several copies.”
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Optimal electroporation parameters for each cell type were determined in preliminary experiments: hDCs (750 V/cm, one pulse of 12 ms); C2C12 (600 V/cm, five pulses of 5 ms within an interval of 400 ms); and HFFs (625 V/cm, one pulse of 24 ms).”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“No statistical methods were applied to pre-determine sample size for animal experiments.”
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The enrichment platform we present here can be adapted with regard to the cell types used for library generation and as a screening environment to find cell type-specific UTR elements for mRNA stabilization, and thus it offers a broadly applicable procedure for the improvement of mRNA therapeutics.”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In summary, AES-mtRNR1- and mtRNR1-AES-based 3′ UTRs, discovered by the approach presented herein, enhance the applications of mRNA-based transfer of genetic information shown exemplarily for antigen-encoding mRNA vaccination and mRNA-based reprogramming of human fibroblasts, and they promise to contribute to clinical translation of mRNA gene delivery.”
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Altogether, these findings indicate a directed and efficient evolution.”
hedged- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In fact, as factors regulating mRNA stability are cell type specific, there may be cell types in which AES-mtRNR1-based elements are not superior over the conventionally used 3′ UTRs.”
hedged- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Rests on an assumption: Factors regulating mRNA stability are cell type specific.
The paper relies on this without support.
mRNA molecules are intrinsically unstable.
“mRNA molecules are intrinsically unstable and their intracellular kinetics depend on the UTRs embracing the coding sequence, in particular the 3′ UTR elements.”
The paper relies on this without support.
hDCs survive in cell culture for an extended period of time without cell division.
“hDCs were chosen as the environment for evolution, as they survive in cell culture for an extended period of time without cell division, which would counteract the enrichment of mRNA species.”
The paper relies on this without support.
hDCs are the host cells for vaccine antigen delivery.
“Moreover, hDCs are the host cells for one of the mRNA applications in which we are interested, namely, vaccine antigen delivery.”
The paper relies on this without support.
Non-integrating, non-self-replicating iPSC generation requires several cycles of transfection with exogenous reprogramming factors until endogenous transcription factors reach sufficiently high protein levels and take over; this interferes with cell viability, increases complexity, and is a hurdle for clinical translation.
“Generation of iPSCs with gene delivery systems that do not integrate and are not self-replicating requires several cycles of transfection with exogenous reprogramming factors until endogenous transcription factors reach sufficiently high protein levels and take over. This interferes with cell viability, increases complexity, and is a hurdle for clinical translation.”
The paper relies on this without support.
Factors regulating mRNA stability are cell type specific.
“as factors regulating mRNA stability are cell type specific”
The paper relies on this without support.
mtRNR1- and AES-derived motives were previously not known to stabilize mRNA.
“mtRNR1- and AES-derived motives were previously not known to stabilize mRNA.”
The paper relies on this without support.
Alphaviral self-replicating RNA comes with the disadvantage that clinical grade iPSCs have to be free of residual contaminations of self-replicating vectors.
“It, however, comes with the disadvantage that clinical grade iPSCs have to be free of residual contaminations of self-replicating vectors.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The cited source doesn’t support the claim as written.
In the citing paper
“As 3′UTRs, various combinations of mtRNR1, AES, and EMCV were chosen, which individually have previously also been found to influence mRNA stability [34,39].”
From Effects of Combinations of Untranslated-Region Sequences on Translation of mRNA (2023) · cites this paper as [34]
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Both of the candidates are pharmacologically optimized 15,16 , N 1 -methylpseudouridine nucleoside-modified mRNAs 17 that are administered intramuscularly in a prime-boost regimen 21”
From BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans (2021) · cites this paper as [16]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“In addition, 5′ UTR: human -globin RNA with optimized Kozak sequence, 3′ UTR: AES and mtRNR1 3′ UTR Motives, and 110 Poly(A) tail with nucleotide-linker (GCAUAUGACU) are also used (Orlandini von Niessen et al. 2019).”
From Modifications of mRNA vaccine structural elements for improving mRNA stability and translation efficiency (2022) · cites this paper as Orlandini von Niessen et al. 2019
Experimental — accuracy not yet measuredThe cited source says what the claim attributes to it.
In the citing paper
“Orlandini von Niessen and coworkers [34] have shown that 3′UTRs containing mtRNR1 and AES in any order promote overexpression of an upstream gene.”
From Effects of Combinations of Untranslated-Region Sequences on Translation of mRNA (2023) · cites this paper as [34]
The check wasn’t confident either way, so it gives no verdict — never disputed.
In the citing paper
“numerous works have developed specific UTRs to increase the translational efficiency of mRNAs, as the use of one or two copies of the 3′ UTR of the globin or other known stable mRNA8,9 or short 14-nucleotide 5′ UTR.27”
From Combining an optimized mRNA template with a double purification process allows strong expression of in vitro transcribed mRNA (2021) · cites this paper as [8]
Experimental — accuracy not yet measured
In the citing paper
“The antigen-encoding RNA contains sequence elements that increase RNA stability and translation efficiency in human dendritic cells 15,16 .”
From BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans (2021) · cites this paper as [16]
In the citing paper
“They showed that 3′UTRs containing mtRNR1 and AES promote the expression of the upstream gene.”
From Untranslated Region Sequences and the Efficacy of mRNA Vaccines against Tuberculosis (2024) · cites this paper as [17]
In the citing paper
“The structural characteristics of mRNA such as the CAP, poly(A) tail, and untranslated regions (UTRs) directly impact mRNA stability and translation efficiency (Orlandini von Niessen et al. 2019; Holtkamp et al. 2006).”
From Modifications of mRNA vaccine structural elements for improving mRNA stability and translation efficiency (2022) · cites this paper as Orlandini von Niessen et al. 2019
In the citing paper
“Of note, despite active research into the properties of UTRs, the number of studies assessing the contribution of certain UTRs to the expression of heterologous RNAs is rather small [34,35].”
From Effects of Combinations of Untranslated-Region Sequences on Translation of mRNA (2023) · cites this paper as [34]
In the citing paper
“BioNTech/Pfizer's modRNA platform RNA sequence (used for BNT162b2) consists of human α-globin 5′ untranslated region (UTR), amino-terminal enhancer of split (AES), and mtRNR1 3′ UTR motifs91 and a poly(A) tail consisting of A30LA70 (linker [L]: GCAUAUGACU),81”
From COVID-19 mRNA vaccines: Platforms and current developments (2022) · cites this paper as [91]
In the citing paper
“BNT162b2 (BioNTech/Pfizer) and mRNA-1273 (Moderna) are composed of 1-methylpseudouridine-modified full-length spike mRNA, with proline substitutions, that is GC rich, codon optimized, and composed of standard mRNA components: cap, 5′ UTR, coding sequence, 3′ UTR, and a poly(A) tail.”
From COVID-19 mRNA vaccines: Platforms and current developments (2022) · cites this paper as [91]
In the citing paper
“However, because UTR performance can vary by cell type, alternative UTR sequences may be used that have been optimized for the desired application and intended cell target16–18.”
From mRNA vaccines for infectious diseases: principles, delivery and clinical translation (2021) · cites this paper as [17]
In the citing paper
“The design of (non-coding) structural elements of the mRNA such as the CAP structure, poly(A) tail and untranslated regions (UTRs) all have a major impact on the mRNA stability and translation capacity [31,32]”
From The dawn of mRNA vaccines: the COVID-19 case (2021) · cites this paper as [31]
Citations
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“Moreover, in line with published data, the length of the 30 UTR elements correlated negatively with their stabilizing effect (Figure S2).(22)”
BackgroundmRNA vaccines — a new era in vaccinology
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Cited by
Sorted by citations
COVID-19 vaccine BNT162b1 elicits human antibody and TH1 T cell responses
Sahin, U., Muik, A., Derhovanessian, E., Vogler, I., Kranz, L. M., Vormehr, M., Baum, A., Pascal, K., Quandt, J., Maurus, D., et al. (2020) · Nature · doi:10.1038/s41586-020-2814-7 · Cited by 1,977
Self-citationmRNA vaccines for infectious diseases: principles, delivery and clinical translation
Chaudhary, N., Weissman, D., & Whitehead, K. A. (2021) · Nature Reviews Drug Discovery · doi:10.1038/s41573-021-00283-5 · Cited by 1,555
Drug delivery systems for RNA therapeutics
Paunovska, K., Loughrey, D., & Dahlman, J. E. (2022) · Nature Reviews Genetics · doi:10.1038/s41576-021-00439-4 · Cited by 1,349
An RNA vaccine drives immunity in checkpoint-inhibitor-treated melanoma
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Self-citationmRNA vaccine for cancer immunotherapy
Miao, L., Zhang, Y., & Huang, L. (2021) · Molecular Cancer · doi:10.1186/s12943-021-01335-5 · Cited by 966
Metadata
- Authors
- Orlandini von Niessen, A. G., Poleganov, M. A., Rechner, C., Plaschke, A., Kranz, L. M., Fesser, S., Diken, M., Löwer, M., Vallazza, B., Beissert, T., Bukur, V., Kuhn, A. N., Türeci, Ö., & Sahin, U.
- Affiliations
- BioNTech (Germany) (DE)
- University Medical Center of the Johannes Gutenberg University Mainz (DE)
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2019-04
- Volume
- 27
- Issue
- 4
- Pages
- 824-836
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC6453560
- OpenAlex
- W2903777694
- Cited by
- 365
- References
- 60
- Citations by year
- 2026: 51 · 2025: 66 · 2024: 68 · 2023: 52 · 2022: 55 · 2021: 45 · 2020: 21 · 2019: 7
- FWCI
- 6.49
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Bundesministerium für Bildung und Forschung
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- synthetic mRNA, mRNA therapeutics, 3' untranslated region, mRNA stability, T cell response, reprogramming factors, induced pluripotency, gene therapy
- MeSH terms
- Animals, Blood Donors, Cells, Cultured, Female, Fibroblasts, Half-Life, Humans, Mice, Inbred BALB C, RNA, Messenger, Vaccination, Genetic Therapy, Gene Library, Gene Transfer Techniques, Cancer Vaccines, 3' Untranslated Regions, RNA Stability, Mice, Induced Pluripotent Stem Cells, Cellular Reprogramming
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