4.1 Article

Single-stranded DNA versus tailed duplex in sequence conversion oflacZαDNA

Journal

NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS
Volume 39, Issue 9, Pages 1245-1250

Publisher

TAYLOR & FRANCIS INC
DOI: 10.1080/15257770.2020.1790596

Keywords

Sequence conversion; gene correction; single-stranded DNA fragment; 5'-tailed duplex; genome editing

Funding

  1. Japan Society for the Promotion of Science (JSPS) KAKENHI grant [JP16H02956]

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Targeted DNA editing has great potential to cure some genetic diseases; however, the use of artificial nucleases such as CRISPR-Cas9 and TALEN in gene therapy can potentially cause severe side effects due to off-target DNA cleavages. Single-stranded (ss) DNAs and 5'-tailed duplexes (TDs) can achieve target base substitutions when introduced without artificial nucleases into cultured cells and mouse liver. In this study, ss DNA and TD were separately co-introduced into human U2OS cells, together with a target plasmid DNA bearing an inactivatedlacZ alpha gene, and the gene correction efficiencies were compared. Unlike the genes examined in previous studies, ss DNA and TD showed similar efficiencies. Therefore, ss DNAs might be as useful as TD for gene correction, depending on the target sequence.

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