RRC ID 65926
Author Suzuki T, Imada T, Komatsu Y, Kamiya H.
Title Comparison of DNA fragments as donor DNAs upon sequence conversion of cleaved target DNA.
Journal Nucleosides Nucleotides Nucleic Acids
Abstract Pinpoint sequence alteration (genome editing) by the combination of the site-specific cleavage of a target DNA and a donor nucleic acid has attracted much attention and the sequence of the target DNA is expected to be changed to that of a donor nucleic acid. In most cases, oligodeoxyribonucleotides (ODNs) and plasmid DNAs have been used as donors. However, a several hundred-base single-stranded (ss) DNA fragment and a 5'-tailed duplex (TD) accomplished the desired sequence changes without DNA cleavage, and might serve as better donors for the cleaved target DNA than ODNs and plasmid DNAs. In this study, sequence conversion efficiencies were compared with various donor DNAs in model sequence alteration experiments, using episomal DNA. The efficiencies with the ss and TD fragments were higher than those with the ODN and plasmid DNA. The sequence change by the TD seemed somewhat less efficient but slightly more accurate than that by the ss DNA fragment. These results suggested that the ss and TD fragments are better donors for targeted sequence alteration.
Volume 36(6)
Pages 428-434
Published 2017-6-3
DOI 10.1080/15257770.2017.1310385
PMID 28486036
MeSH Base Sequence DNA Cleavage* DNA, Single-Stranded / chemistry* DNA, Single-Stranded / genetics*
IF 1.167
Human and Animal Cells HeLa(RCB0007)