Extended Data Table 2. Off-target sites identified by CIRCLE-seq for gP, gM, and gMH that exhibit single nucleotide polymorphisms (SNPs) based on CIRCLE-seq data.

Mismatches relative to the on-target site are shown as lower-case letters. SNPs that differ from the C57BL/6N mouse strain are shown as red-colored lowercase letters. Targeted amplicon sequencing of sites for gM (first row) was performed with the same genomic DNA from WT and KI mice used for CIRCLE-seq experiments (data in Supplementary Table 7). N.D. = not done.

Location of site	gRNA	Mouse	gRNA on-target site	Off-target sequence	Number of mismatches	SNPs confirmed by targeted amplicon- sequencing
chr9:78832014-78832037	gM	KI	GGCT GAT GAGGCCGCACATGNGG	atCAGAT aAaaCCaCACATGGag	8	No
chr4:129226173-129226196	gMH	KI	CAGGTTCCATGGGATGCTCTNGG	agGGcT CacctGGATGCTCTGtG	8	N.D.
chr9:68916733-68916756	gMH	KI	CAGGTTCCATGGGATGCTCTNGG	tAGGgagagaGGATGCTCTGaG	8	N.D.
chr3:19461541-19461564	gMH	KI	CAGGTTCCATGGGATGCTCTNGG	CAtGTaCCAaGGGATGtTCTAcG	5	N.D.
chr13:37109353-37109376	gP	KI	AGCAGCAGCGGCGGCAACAGNGG	a-cAGCAGCAGCAGCAACAACGA	6	N.D.
chr6:112201818-112201841	gp	WT	AGCAGCAGCGGCGGCAACAGNGG	AGCAAcagcagcagcagcagtAG	5	N.D.