GuideSeq

GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases

GUIDE-Seq (Genome-wide, Unbiased Identification of DSBs Enabled by Sequencing) is a molecular biology technique that allows for the unbiased in vitro detection of off-target genome editing events in DNA caused by CRISPR/Cas9 as well as other RNA-guided nucleases (RGN) in living cells. Similar to LAM-PCR, it employs multiple PCRs to amplify regions of interest that contain a specific insert that preferentially integrates into double-stranded breaks. As gene therapy is an emerging field, GUIDE-Seq has gained traction as a cheap method to detect the off-target effects of potential therapeutics without needing whole genome sequencing.