Details

Autor(en) / Beteiligte

• Nobles, Christopher L
• Reddy, Shantan
• Salas-McKee, January
• Liu, Xiaojun
• June, Carl H
• Melenhorst, J Joseph
• Davis, Megan M
• Zhao, Yangbing
• Bushman, Frederic D

Titel

iGUIDE: an improved pipeline for analyzing CRISPR cleavage specificity

Ist Teil von

• Genome Biology, 2019-01, Vol.20 (1), p.14-14, Article 14

Ort / Verlag

England: BioMed Central

Erscheinungsjahr

2019

Link zum Volltext

Quelle

SpringerNature Journals

Beschreibungen/Notizen

• 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 .