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High-Throughput 5’ UTR Engineering for Enhanced Protein Production in Non-Viral Gene Therapies

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE176581 | GEO | 2021/06/11

REPOSITORIES: GEO

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