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Cited by 117RNA Interference and Gene DeliveryRNA and protein synthesis mechanismsNanoparticle-Based Drug Delivery

Single-cell mRNA transfection studies: Delivery, kinetics and statistics by numbers

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Leonhardt, Carolin, Schwake, Gerlinde, Stögbauer, Tobias R., Rappl, Susanne, Kuhr, Jan-Timm, Ligon, Thomas S., Rädler, Joachim O.·2014·Nanomedicine: Nanotechnology, Biology and Medicine
DOI 10.1016/j.nano.2013.11.008
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Abstract

In artificial gene delivery, messenger RNA (mRNA) is an attractive alternative to plasmid DNA (pDNA) since it does not require transfer into the cell nucleus. Here we show that, unlike for pDNA transfection, the delivery statistics and dynamics of mRNA-mediated expression are generic and predictable in terms of mathematical modeling. We measured the single-cell expression time-courses and levels of enhanced green fluorescent protein (eGFP) using time-lapse microscopy and flow cytometry (FC). The single-cell analysis provides direct access to the distribution of onset times, life times and expression rates of mRNA and eGFP. We introduce a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby the dose-response relation. Our results establish a statistical framework for mRNA transfection and as such should advance the development of RNA carriers and small interfering/micro RNA-based drugs. FROM THE CLINICAL EDITOR: This team of authors established a statistical framework for mRNA transfection by using a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby their dose-response relation. This study establishes a nice connection between theory and experimental planning and will aid the cellular delivery of mRNA molecules.

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  • Chemical structure mapping showed that the long-lived LinearDesign-1 was significantly more highly structured than Yellowstone, as assessed by DMS and SHAPE reactivities.

    “Chemical structure mapping showed that the long-lived LinearDesign-1 was significantly more highly structured than Yellowstone, as assessed by dimethyl sulfate (DMS) and selective 2’-hydroxyl acylation with primer extension (SHAPE) reactivities59,60 (Fig. 2H, Fig. S3) and structure models guided by these data (Fig. 2I).”

    From Combinatorial optimization of mRNA structure, stability, and translation for RNA-based therapeutics (2022) · cites this paper as [59]

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Authors
Leonhardt, C., Schwake, G., Stögbauer, T. R., Rappl, S., Kuhr, J.-T., Ligon, T. S., & Rädler, J. O.
Affiliations
  • Center for NanoScience (DE)
  • Technische Universität Berlin (DE)
Corresponding author
Rädler, Joachim O.
ORCID
Venue
Nanomedicine: Nanotechnology, Biology and Medicine
Publisher
Elsevier BV
Published
2014-05
Volume
10
Issue
4
Pages
679-688
ISSN
1549-9634 (ISSN-L)
DOI
10.1016/j.nano.2013.11.008
OpenAlex
W1974595716
Cited by
117
References
3
Citations by year
2026: 8 · 2025: 10 · 2024: 9 · 2023: 10 · 2022: 19 · 2021: 9 · 2020: 11 · 2019: 11 · 2018: 9 · 2017: 6 · 2016: 8 · 2015: 5 · 2014: 2
FWCI
1.88
Type
journal article
Language
en
Open access
Licence
Locations
  • Nanomedicine Nanotechnology Biology and Medicine (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
  • Deutsche Forschungsgemeinschaft (SFB1032,)
  • Elitenetzwerk Bayern
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 and protein synthesis mechanisms
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
mRNA transfection, single-cell gene expression, enhanced green fluorescent protein, time-lapse microscopy, flow cytometry, dose-response relationship, mRNA translation
MeSH terms
Humans, Kinetics, RNA, Messenger, Transfection, Cell Line, Tumor, Green Fluorescent Proteins

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