iGUIDE: an improved pipeline for analyzing CRISPR cleavage specificity

摘要: Genome engineering methods have advanced greatly with the development of programmable nucleases, but for quantifying on- and off-target cleavage sites associated deletions remain nascent. Here, we report an improvement GUIDE-seq method, iGUIDE, which allows filtering mispriming events to clarify true signal. Using specify locations Cas9-guided four guide RNAs, characterize deletions, show that naturally occurring background DNA double-strand breaks are open chromatin, gene dense regions, chromosomal fragile sites. iGUIDE is available from https://github.com/cnobles/iGUIDE .