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Table 2 Properties of engineered CRRISPR-Cas9 variants

From: CRISPR-Cas gene editing technology and its application prospect in medicinal plants

Engineered Cas9 variants Included mutations PAM (5′ to 3′) Notes Refs.
SpCas9 VRER D1135V, G1218R, R1335E, T1337R of SpCas9 mutations NGCG Altered PAM variant; Bacterial-selection-based screening [41, 60]
SpCas9 VQR D1135V, R1335Q, T1337R of SpCas9 mutations NGAN or NGNG Altered PAM variant; Bacterial-selection-based screening [41, 60]
SpCas9 EQR D1135E, R1335Q, T1337R of SpCas9 mutations NGAG Altered PAM variant; Bacterial-selection-based screening [61]
SpCas9-NG R1335V, L1111R, D1135V, G1218R, E1219F, A1322R, T1337R of SpCas9 mutations NG Altered PAM variant [62]
SpG D1135L, S1136W, G1218K, E1219Q, R1335Q, T1337R of SpCas9 mutations NGN A near-PAMless variant [43, 63]
SpRY A61R, L1111R, N1317R, A1322R, R1333P introduced into SpG NRN, NYN A near-PAMless variant [43, 63]
xCas9 3.7 E480K, E543D, E1219V, A262T, R324L, S409I, M694I of SpCas9 mutations NG, GAA, GAT Expanded PAM recognition range; Phage-assisted continuous evolution (PACE) [25, 59]
SpCas9-HF1 N497A, R661A, Q695A, Q926A of SpCas9 mutations NGG Enhanced specificity [64]
eSpCas9 (1.0) K810A, K1003A, R1060A of SpCas9 mutations NGG Enhanced specificity; Structure-guided protein engineering [65]
eSpCas9 (1.1) K848A, K1003A, R1060A of SpCas9 mutations NGG Enhanced specificity; Structure-guided protein engineering [65, 66]
evoCas9 M495V, Y515N, K526E, R661Q of SpCas9 mutations NGG Enhanced specificity; Yeast-based screening [42]
HypaCas9 N692A, M694A, Q695A, H698A of SpCas9 mutations NGG Enhanced specificity [67]
HiFi Cas9 single point mutation R691A of SpCas9 NGG Enhanced specificity for ribonucleoprotein delivery [68]
KKH SaCas9 E782K, N968K, R1015H of SaCas9 mutations NNNRRT Altered PAM variant [69, 70]
SaCas-HF R245A, N413A, N419A, R654A of SaCas9 mutations NNGRRT Enhanced specificity and genome-wide targeting accuracy [69, 70]
efSaCas9 single point mutation N260D of SaCas9 variant Mut268 NNGRRT Enhanced specificity; Human cells-based screening [71, 72]
(HiFi-)Sc++ Thr1227Lys, Arg701Ala mutations and loop sequence from S. anginosus introduced into ScCas9 NNG Enhanced specificity and activity [73, 74]