4.4 Article

Correction of monomeric enhanced green fluorescent protein (mEGFP) gene by short 5'-tailed duplexes

Journal

JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 134, Issue 3, Pages 175-181

Publisher

SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2022.06.014

Keywords

Gene correction; Gene editing; 5'-tailed duplex; mEGFP gene; copGFP

Funding

  1. Japan Society for the Promotion of Science (JSPS) KAKENHI [JP 17K19491]

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Mutations of important genes can cause various disorders, including cancer. Recent research has shown that a new version of a double-stranded structure can correct base substitution mutations in target genes in human cells. The experiments demonstrated the effective gene editing capabilities of this structure.
Mutations of important genes elicit various disorders, including cancer. Recently, a new version of a 5'-tailed duplex (short TD), consisting of a -100-base editor strand containing the wild-type sequence and a -35-base assistant strand, was shown to correct a base substitution mutation in a target gene in human cells. In that previous study, the target was the copepod green fluorescent protein (copGFP) gene. To examine the usefulness of the short TD, we performed gene correction experiments using a mutant form of the monomeric enhanced Aequorea victoria green fluorescent protein (mEGFP) gene containing a TAC to CAC mutation in codon 75 (corresponding to the tyrosine to histidine substitution in the chromophore). The short TDs with the wild-type sequence efficiently corrected the inactivated gene in human U2OS cells. These results indicated that the short TDs are effective for gene editing. (c) 2022, The Society for Biotechnology, Japan. All rights reserved.

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