Table 2 On-target and off-target gene-editing activity by deep sequencing.

	HEK293T		HEK293T		HEK293T		HEK293T		HEK293T
	CR1		CR3		CR5		CR6		CR36
	% Indels	Treated/untreated	% Indels	Treated/untreated	% Indels	Treated/untreated	% Indels	Treated/untreated	% Indels	Treated/untreated
ON	43.347	833.596	47.12	889.13	49.82	503.20	41.74	1391.40	46.65	1227.66
OT1	2.714	100.519	14.21	1092.92	0.056	1.24	0.009	1.29	0.047	1.12
OT2	0.203	9.227	0.018	1.20	0.007	0.70	0.022	1.38	NA	NA
OT3	0.020	1.250	0.026	0.84	0.007	1.75	0.024	1.04	1.179	30.23
OT4	0.051	0.927	0.013	0.81	0.016	0.89	0.014	0.70	0.174	1.09
OT5	0.175	6.250	0.012	0.44	0.01	0.71	NA	NA	0.138	1.12
OT6	0.057	1.295	0.038	1.81	0.064	3.05	0.141	1.10	0.062	1.19
OT7	0.117	2.294	0.032	1.07	0.019	1.00	0.116	1.20	0.02	0.87
OT8	0.068	1.478	0.011	0.58	0.127	1.40	0.008	1.60	0.215	0.67
OT9	0.089	0.967	0.014	0.70	0.018	1.06	0.042	1.24	0.014	0.78
OT10	0.030	0.612	0.01	0.53	0.023	1.10	0.013	0.68	0.21	6.77

DMD, Duchenne muscular dystrophy; NA, not applicable.
HEK293Ts were transfected with constructs encoding human codon-optimized SpCas9 and the indicated sgRNA. The frequency of indel formation at each target site (ON, bold) and the top ten predicted off-target sites (OT1–10) was determined by deep sequencing. Two off-target sites (CR6-OT5 and CR36-OT2) did not produce reads that met filtering criteria. Off-target sites with activity greater than 0.2% or greater than tenfold increase compared with untreated controls are highlighted in red. These off-target sites and their corresponding on-target site were also assessed in the treated, sorted DMD myoblasts (bottom).

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