Genome engineering methods have advanced greatly with the development of programmable nucleases, but methods for quantifying on- and off-target cleavage sites and associated deletions remain nascent. Here, we report an improvement of the GUIDE-seq method, iGUIDE, which allows filtering of mispriming events to clarify the true cleavage signal. Using iGUIDE, we specify the locations of Cas9-guided cleavage for four guide RNAs, characterize associated deletions, and show that naturally occurring background DNA double-strand breaks are associated with open chromatin, gene dense regions, and chromosomal fragile sites. iGUIDE is available from https://github.com/cnobles/iGUIDE.
CITATION STYLE
Nobles, C. L., Reddy, S., Salas-Mckee, J., Liu, X., June, C. H., Melenhorst, J. J., … Bushman, F. D. (2019). IGUIDE: An improved pipeline for analyzing CRISPR cleavage specificity Jin-Soo Kim. Genome Biology, 20(1). https://doi.org/10.1186/s13059-019-1625-3
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