gRNA validation for wheat genome editing with the CRISPR-Cas9 system

BMC Biotechnology

Table 1 Experimentally determined versus in silico predicted on-target gRNA activity

	Target homoeoalleles	Indel frequency (%)^a	sgRNA Designer score^b	WU-CRISPR score^c
gRNA1	7AS	1.3	0.59	58
gRNA2	7AS	9.8	0.58	60
gRNA3	7AS, 4AL, 7DS	1.8	0.64	82
gRNA4	7AS, 4AL, 7DS	7.0	0.52	< 50
gRNA5	7AS, 4AL, 7DS	17.8	0.68	85
gRNA6	7AS, 4AL, 7DS	0.0	0.47	< 50
gRNA7	7AS, 4AL	2.5	0.85	< 50

^aExperimentally determined on-target gRNA activity, expressed as the mean proportion of edited amplicon reads derived from target homoeoalleles for three replicates. ^bPotential range = 0–1 (1 is highest predicted gRNA activity); c potential range = 0–100 (100 is highest predicted gRNA activity, scores < 50 are not output)

ISSN: 1472-6750