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https://doi.org/10.1371/journal.pone.0082100
| Title: | An internal ribosome entry site (IRES) mutant library for tuning expression level of multiple genes in mammalian cells | Authors: | Koh E.Y.C. Ho S.C.L. Mariati Bioprocessing Technology Institute Song Z. Bi X. Bardor M. Yang Y. |
Keywords: | monoclonal antibody myosin heavy chain myosin light chain immunoglobulin heavy chain immunoglobulin light chain animal cell article codon controlled study Encephalomyocarditis virus enzyme linked immunosorbent assay gene construct gene expression gene mutation gene vector human human cell internal ribosome entry site liquid chromatography mammal cell mass spectrometry nonhuman open reading frame polyacrylamide gel electrophoresis protein cleavage start codon supernatant translation initiation Western blotting animal cell line gel chromatography gene expression regulation genetic transfection genetics mammal metabolism mutation nucleotide sequence ribosome Animals Base Sequence Blotting, Western Cell Line Chromatography, Gel Encephalomyocarditis virus Gene Expression Regulation Humans Immunoglobulin Heavy Chains Immunoglobulin Light Chains Mammals Mutation Ribosomes Transfection Cell Line Gene Expression Gene Order Humans Open Reading Frames Recombinant Fusion Proteins Repetitive Sequences, Nucleic Acid RNA RNA, Long Noncoding |
Issue Date: | 2013 | Citation: | Koh E.Y.C., Ho S.C.L., Mariati Bioprocessing Technology Institute, Song Z., Bi X., Bardor M., Yang Y. (2013). An internal ribosome entry site (IRES) mutant library for tuning expression level of multiple genes in mammalian cells. PLoS ONE 8 (12) : e82100. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0082100 | Rights: | Attribution 4.0 International | 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 10th, 11th, 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. © 2013 Koh et al. | Source Title: | PLoS ONE | URI: | https://scholarbank.nus.edu.sg/handle/10635/161448 | ISSN: | 1932-6203 | DOI: | 10.1371/journal.pone.0082100 | Rights: | Attribution 4.0 International |
| Appears in Collections: | Staff Publications Elements |
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