View Full Table | Close Full ViewTable 1.

Plant CRISPR studies where off-target mutations were analyzed.

 
Species Cas9 codon optimization Cas9 promoter gRNA promoter Off-target effect Generation monitored† Off-target analysis‡ Reference
Transient transfection
Arabidopsis thaliana Arabidopsis 35SPPDK AtU6 No NA No mutation was detected in a near-homolog to the target containing two mismatches to the 12-bp seed sequence Li et al., 2013
Citrus sinensis Human CaMV35S CaMV35S No NA No mutation for eight sites with 13- to 17-bp out of 20-bp identity to the target sequence Jia and Wang, 2014
Nicotiana benthamiana Eukaryotic CaMV35S AtU6 No NA No mutation for 18 sites with 14- to 17-bp out of 20-bp identity to the target sequence Nekrasov et al., 2013
Oryza sativa Rice 2xCaMV35S OsU3 Yes NA Mutations occurring at one of three near-homologous sites with 1- or 3-bp mismatches to the target sequences Shan et al., 2013
Oryza sativa Human CaMV35S OsU3 or OsU6 Yes NA Mutations occurred at one of three mismatch sites containing one mismatch in the 15-bp seed sequence; mutation occurred for a mismatch at the 11 position proximal to the PAM but not at the 1 or 9 position Xie and Yang, 2013
Stable transformation
Arabodopsis thaliana Human CaMV35S AtU6 No T1, T2 Whole-genome sequencing of T1 and T2 plants at a sequencing depth of ∼60x Feng et al., 2014
Glycine max Human CaMV35S MtU6 Yes T0 Three of 15 potential off-target sites with two to six mismatches were edited in a range of frequencies Jacobs et al., 2015
Nicotiana tabacum Tobacco 2xCaMV35S AtU6 No T0 No mutation for one potential off-target site with a perfect match to the 12-bp seed sequence and having 18-bp out of 20-bp identity with the target sequence Gao et al., 2015
Oryza sativa Rice 2xCaMV35S ZmU3 No T0 Whole-genome sequencing of seven T0 plants Zhang et al., 2014
Oryza sativa Rice 2xCaMV35S ZmU3 Yes T0 One mutation in analysis of 13 putative mismatch loci containing from one to six mismatched bases; the susceptible loci had a 1-bp mismatch outside the seed region of the target site and showed mutation for seven of 72 T0 plants Zhang et al., 2014
Oryza sativa Plant 2xCaMV35S AtU6 No T0 No mutation detected for three near-homologous sites with 1-base or 3-base mismatches to the target sequence Xu et al., 2014
Oryza sativa Rice ZmUbi OsU6 No T0 No mutations occurred at six sites that contained 16 or more nucleotide matching the target sequence Zhou et al., 2014
Oryza sativa Rice 2xCaMV35S OsU3 Yes T0 Frequent mutations of a homologous sequence with a 1-bp mismatch 17-bp 5′ upstream of the PAM sequence Endo et al., 2014
Populus trichocarpa Human CaMv35S MtU6 No T0 No mutation in closest homolog with four mismatches including one in PAM; no mutation in transgenic Zhou et al., 2014
Zea maize Maize ZmUbi ZmU6 No T0, T1 No mutations for eight sites with five or fewer mismatches to the target sequence Zhu et al., 2016
NA, not applicable.
PAM, protospacer adjacent motif.