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Table 1 Summary of reported on- and off-target mutation frequencies of single and dual CRISPR-Cas9 systems

From: RNA-guided CRISPR-Cas technologies for genome-scale investigation of disease processes

Investigators Platform Reported on-targeting (%) Reported off-targeting (%)
Shalem et al., 2014 [39] Single Cas platform ~100 0–15b
Wang et al., 2014 [41] Single Cas platform 97 0–29.5c
Koike-Yusa et al., 2014 [42] Single Cas platform 12.7 ~0.7d
Tsai et al., 2014 [46] Dual Cas platform 3–40 0
Ran et al., 2013 [48] Dual Cas platform 40a 0
Guilinger et al., 2014 [47] Dual Cas platform 8–22 0
  1. aOn-target mutation frequencies of up to 40% were observed in target sequences when the distance between the Cas9 nickase pairs were between −4 and +20 bp.
  2. b9 of 12 sgRNAs showed minimal (~0–3%) modification of predicted off-target sites. 3 of 12 sgRNAs showed modification (~15% or higher) on at least 1 of their off-target sites.
  3. c0–2.5% cleavage of 12 of the 13 predicted off-target sites, 29.5% cleavage of 1 of the 13 predicted off-target sites.
  4. dCleavage analysis of 275 potential off-target sites for the Pigga site 2 sgRNA revealed only 2 of those potential off-target sites were cleaved and both were in noncoding regions (2/275*100 = ~0.7%).