An Internal Ribosome Entry Site (IRES) Mutant Library for Tuning Expression Level of Multiple Genes in Mammalian Cells

Esther Koh (1) , Steven Ho (1) , . Mariati (1) , Zhiwei Song (1) , Xuezhi Bi (1) , Muriel Bardor (2, 1) , Yuansheng Yang (1) , Linda Hendershot (1, 3)

1 A*STAR - Agency for science, technology and research [Singapore]
2 Glyco-MEV - Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale
3 Nanyang Technological University [Singapour]

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.

Domains: Chemical Sciences; Polymers; Life Sciences [q-bio]; Vegetal Biology; Plant breeding; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Biochemistry [q-bio.BM]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular biology; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Genomics [q-bio.GN]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular Networks [q-bio.MN]; Life Sciences [q-bio]; Vegetal Biology; Phytopathology and phytopharmacy; Life Sciences [q-bio]; Biotechnology; Life Sciences [q-bio]; Development Biology; Gametogenesis; Life Sciences [q-bio]; Cellular Biology; Subcellular Processes [q-bio.SC]; Life Sciences [q-bio]; Cellular Biology; Cell Behavior [q-bio.CB]

Esther Koh, Steven Ho, . Mariati, Zhiwei Song, Xuezhi Bi, et al.. An Internal Ribosome Entry Site (IRES) Mutant Library for Tuning Expression Level of Multiple Genes in Mammalian Cells. PLoS ONE, 2013, 8 (12), pp.e82100. ⟨10.1371/journal.pone.0082100⟩. ⟨hal-01843928⟩