title: An internal ribosome entry site (IRES) mutant library for tuning expression level of multiple genes in mammalian cells.
abstract: 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.
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
authors.forename: Esther Y C
authors.initials: EY
authors.lastname: Koh
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
authors.forename: Steven C L
authors.initials: SC
authors.lastname: Ho
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
authors.lastname: Mariati
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Song
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Bi
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore ; Laboratoire Glyco-MEV (Glycobiologie et Matrice extracellulaire végétale), Université de Rouen, Mont-Saint-Aignan, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Bardor
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore ; School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Yuansheng
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002850:Chromatography, Gel; D004686:Encephalomyocarditis virus / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D008322:Mammals / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism*; D014162:Transfection
chemical_list: D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains