Characterization of DSB repair pathway choice with biased genome editing

We analyzed the effect of CtIP overexpression and CtIP accumulation at DSB site by LoAD system in terms of the accuracy and repair pathway choice. The short 3? Flag tag was knocked-in in this assay to enable sequence analysis covering the full length (left microhomology, knock-in fragment, and right microhomology) by next-generation sequencing (NGS). For the unbiased analysis of all analyzable alleles including precise knock-in, imprecise knock-in, and non-knock-in ones, we performed out-out PCR of genomic integration sites at the ATP5B, PARP1, and RPL11 loci, followed by NGS analysis.