High-throughput 5′ UTR engineering for enhanced protein production in non-viral gene therapies
| العنوان: | High-throughput 5′ UTR engineering for enhanced protein production in non-viral gene therapies |
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| المؤلفون: | Gigi C G Choi, Eva Maria Novoa, Claudia Wehrspaun, Alan S.L. Wong, Timothy K. Lu, William C.W. Chen, Jicong Cao, Zhizhuo Zhang, Dianbo Liu, Manolis Kellis |
| المصدر: | Nature Communications Nature Communications, Vol 12, Iss 1, Pp 1-10 (2021) |
| بيانات النشر: | Nature Publishing Group UK, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0301 basic medicine, Untranslated region, Translation, Five prime untranslated region, Genetic enhancement, Science, In silico, General Physics and Astronomy, Gene Expression, Computational biology, Biology, General Biochemistry, Genetics and Molecular Biology, Article, Cell Line, Recombinases, 03 medical and health sciences, Synthetic biology, Plasmid, 0302 clinical medicine, Gene expression analysis, Gene expression, Protein biosynthesis, Recombinase, Humans, Promoter Regions, Genetic, Gene, 030304 developmental biology, 0303 health sciences, Multidisciplinary, HEK 293 cells, High-throughput screening, Translation (biology), General Chemistry, Genetic Therapy, 3. Good health, High-Throughput Screening Assays, ComputingMethodologies_PATTERNRECOGNITION, 030104 developmental biology, HEK293 Cells, 030220 oncology & carcinogenesis, Teràpia genètica, 5' Untranslated Regions, Genetic Engineering, Proteïnes, Genètica, Algorithms, Plasmids |
| الوصف: | Despite significant clinical progress in cell and gene therapies, maximizing protein expression in order to enhance potency remains a major technical challenge. Here, we develop a high-throughput strategy to design, screen, and optimize 5′ UTRs that enhance protein expression from a strong human cytomegalovirus (CMV) promoter. We first identify naturally occurring 5′ UTRs with high translation efficiencies and use this information with in silico genetic algorithms to generate synthetic 5′ UTRs. A total of ~12,000 5′ UTRs are then screened using a recombinase-mediated integration strategy that greatly enhances the sensitivity of high-throughput screens by eliminating copy number and position effects that limit lentiviral approaches. Using this approach, we identify three synthetic 5′ UTRs that outperform commonly used non-viral gene therapy plasmids in expressing protein payloads. In summary, we demonstrate that high-throughput screening of 5′ UTR libraries with recombinase-mediated integration can identify genetic elements that enhance protein expression, which should have numerous applications for engineered cell and gene therapies. The engineering of 5′ UTRs that modulate protein expression remains a great challenge. Here we leverage synthetic biology and computational design to develop a high-throughput strategy to design, screen, and optimize 5′ UTRs that enhance protein expression for non-viral gene therapies. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 2041-1723 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d7bf813d08f2e99f07aa027c2756cec2 http://europepmc.org/articles/PMC8260622 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....d7bf813d08f2e99f07aa027c2756cec2 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 20411723 |
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