Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells - mcdbucsb
). An intriguing possibility is therefore that xHDRT ICLs are uncovered during recombination between the DSB and the template. Validation of such a model in the context of our observation that crosslinks stimulate xHDRT recombination inwould suggest that HDR is explored frequently during DSB repair and that detection of crosslinked DNA increases the likelihood that HDR will proceed.
All dsDNAs were derived from purified plasmid DNA from bacterial cultures containing the indicated plasmid or by SPRI purification of amplified linear dsDNA.in 1× TE buffer in a 1.5 ml microcentrifuge tube. Psoralen was then added to the reaction tube to the desired final concentration. Each reaction mixture in an open microcentrifuge tube, placed on ice, was then irradiated with long wavelength UV for 15 min in a Spectrolinker XL-1000 at 365 nm.
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