An Internal Ribosome Entry Site (IRES) Mutant Library for Tuning Expression Level of Multiple Genes in Mammalian Cells
Résumé
A set of mutated Encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) elements with varying strengths is generated by mutating the translation initiation codons of 10 th , 11 th , and 12 th AUG to non-AUG triplets. They are able to control the relative expression of multiple genes over a wide range in mammalian cells in both transient and stable transfections. The relative strength of each IRES mutant remains similar in different mammalian cell lines and is not gene specific. The expressed proteins have correct molecular weights. Optimization of light chain over heavy chain expression by these IRES mutants enhances monoclonal antibody expression level and quality in stable transfections. Uses of this set of IRES mutants can be extended to other applications such as synthetic biology, investigating interactions between proteins and its complexes, cell engineering, multi-subunit protein production, gene therapy, and reprogramming of somatic cells into stem cells.
Domaines
Polymères Amélioration des plantes Biochimie [q-bio.BM] Biologie moléculaire Génomique, Transcriptomique et Protéomique [q-bio.GN] Réseaux moléculaires [q-bio.MN] Phytopathologie et phytopharmacie Biotechnologies Gamétogenèse Organisation et fonctions cellulaires [q-bio.SC] Interactions cellulaires [q-bio.CB]
Origine : Publication financée par une institution
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