Please use this identifier to cite or link to this item: 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
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