IGUIDE: an Improved Pipeline for Analyzing CRISPR Cleavage Specificity

Overview

Journal Genome Biol

Specialties Biology
Genetics

Date 2019 Jan 19

PMID 30654827

Citations 38

Authors

Christopher L Nobles

Shantan Reddy

January Salas-Mckee

Xiaojun Liu

Carl H June

J Joseph Melenhorst

Megan M Davis

Yangbing Zhao

Frederic D Bushman

Affiliations

Soon will be listed here.

Abstract

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 .

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