期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 132, 期 6, 页码 552-559出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2021.08.012
关键词
Gene correction; Gene editing; 5 '-tailed duplex; Single-stranded DNA; copGFP gene
资金
- Japan Society for the Promotion of Science KAKENHI [JP 17K19491]
The study found that 5'-TDs with 100-base ss editor DNAs were as effective in gene correction as those with longer editor DNAs. Additionally, the antisense strand was more effective as the editor than the sense strand. These results suggest that 5'-TD fragments with shorter editor strands are useful for gene correction.
September Various diseases, including cancer, are caused by genetic mutations. A 5'-tailed duplex (TD) DNA, consisting of a long single-stranded (ss) editor DNA and a short (35-base) ss assistant oligodeoxyribonucleotide, can introduce a base substitution in living cells and thus correct mutated genes. Previously, several hundred-base DNAs were employed as the editor DNAs. In this study, 5'-TDs were prepared from various editor DNAs with different lengths and examined for their gene correction abilities, using plasmid DNA bearing a mutated copepod green fluorescent protein (copGFP) gene, in human cells. High-throughput analysis was performed by the reactivated fluorescence of the wild-type protein encoded by the corrected gene as the indicator. The analysis revealed that 5'-TDs with 100-base ss editor DNAs enabled gene editing at least as efficiently as those with longer editor DNAs. Moreover, the antisense strand was more effective as the editor than the sense strand, in contrast to the 5'-TDs with longer editor strands. These results indicated that the 5'-TD fragments with shorter editor strands than those used in previous studies are useful nucleic acids for gene correction. (C) 2021, The Society for Biotechnology, Japan. All rights reserved.
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