Human cellular CYBA UTR sequences increase mRNA translation without affecting the half-life of recombinant RNA transcripts
Open accessAbstract
Modified nucleotide chemistries that increase the half-life (T 1/2) of transfected recombinant mRNA and the use of non-native 5′- and 3′-untranslated region (UTR) sequences that enhance protein translation are advancing the prospects of transcript therapy. To this end, a set of UTR sequences that are present in mRNAs with long cellular T 1/2 were synthesized and cloned as five different recombinant sequence set combinations as upstream 5′-UTR and/or downstream 3′-UTR regions flanking a reporter gene. Initial screening in two different cell systems in vitro revealed that cytochrome b-245 alpha chain (CYBA) combinations performed the best among all other UTR combinations and were characterized in detail. The presence or absence of CYBA UTRs had no impact on the mRNA stability of transfected mRNAs, but appeared to enhance the productivity of transfected transcripts based on the measurement of mRNA and protein levels in cells. When CYBA UTRs were fused to human bone morphogenetic protein 2 (hBMP2) coding sequence, the recombinant mRNA transcripts upon transfection produced higher levels of protein as compared to control transcripts. Moreover, transfection of human adipose mesenchymal stem cells with recombinant hBMP2-CYBA UTR transcripts induced bone differentiation demonstrating the osteogenic and therapeutic potential for transcript therapy based on hybrid UTR designs.
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- Authors
- Ferizi, M., Aneja, M. K., Balmayor, E. R., Badieyan, Z. S., Mykhaylyk, O., Rudolph, C., & Plank, C.
- Affiliations
- TUM Klinikum (DE)
- Ethris (Germany) (DE)
- Corresponding author
- Balmayor, Elizabeth R.
- Plank, Christian
- Venue
- Scientific Reports
- Published
- 2016-12-15
- Volume
- 6
- Issue
- 1
- Pages
- 39149-39149
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC5156912
- OpenAlex
- W2562374902
- Semantic Scholar
- d5079010e9878ffd2688df78e422f62798b4a113
- Cited by
- 46
- References
- 51
- Citations by year
- 2026: 5 · 2025: 12 · 2024: 3 · 2023: 5 · 2022: 5 · 2021: 5 · 2020: 2 · 2019: 3 · 2018: 6
- FWCI
- 1.18
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- mediaTUM – the media and publications repository of the Technical University Munich (Technical University Munich) (submittedVersion, other-oa)
- mediaTUM – the media and publications repository of the Technical University Munich (Technical University Munich) (submittedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Bundesministerium für Bildung und Forschung (01GM1106E, 0315986)
- Technische Universität München
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- 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
- Keywords
- 5' untranslated region, 3' untranslated region, CYBA, mRNA translation, mRNA stability, modified nucleotides, BMP-2, adipose-derived stem cells, osteogenic differentiation, reporter genes
- MeSH terms
- A549 Cells, Adipose Tissue, Animals, Half-Life, Humans, Osteogenesis, Recombinant Proteins, RNA, Messenger, ROC Curve, Transfection, Protein Biosynthesis, Genes, Reporter, NADPH Oxidases, Area Under Curve, 5' Untranslated Regions, 3' Untranslated Regions, RNA Stability, NIH 3T3 Cells, Mice, Bone Morphogenetic Protein 2, Mesenchymal Stem Cells
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