Table 1.
Overview of CRISPRa and CRISPRi platforms
| Cas variant | PAM | Effector/recruiters | Total size (bp) | Reference |
|---|---|---|---|---|
| dSpCas9 | 5′-NGG-3′ | – | ∼4,101 | Qi et al.; Jinek et al.2,13 |
| KRAB | ∼4,341 | Jensen et al.; Tian et al.; le Sage et al.; Dräger et al.; Jung et al.; Schmidt et al.; Cui et al.; Drobna-Śledzińska et al.; Chung et al.; Heman-Ackah et al.; Moreno et al.; Yoshida et al.; Escobar et al.; Leng et al.26,27,28,29,30,31,32,33,34,35,36,37,38,39 | ||
| KRAB-MeCP2 | ∼5,313 | Yeo et al.; Alerasool et al.; de Solis et al.; Ding et al.40,41,42,43 | ||
| ZIM3 | ∼4,599 | Alerasool et al.41 | ||
| VP64a | ∼4,380 | Ding et al.; Matharu et al.; Black et al.; Sokka et al.; Zhou et al.; Colasante et al.; Zhu et al.; Thompson et al.; Jaudon et al.43,44,45,46,47,48,49,50,51 | ||
| VPR | ∼5,799 | Jensen et al.; Cui et al.; Heman-Ackah et al.; Escobar et al.; Strezoska et al.; Urrutia-Cabrera et al.; Nguyen et al.; Unlu et al.; Susco et al.; Schoger et al.; Di Maria et al.26,32,35,38,52,53,54,55,56,57,58 | ||
| SunTag (GCN4-VP64) | ∼4,998 + ∼1,749b | Jung et al.; Tanenbaum et al.; Liu et al.; Boettcher et al.; Liu et al.; Danziger et al.; Siepe et al.30,59,60,61,62,63,64 | ||
| SunTag (GCN4-p65-HSF1) | ∼5,157 + ∼1,929b | Zhou et al.65 | ||
| VP64 (MS2-p65-HSF1) (SAM) | ∼4,380 + ∼1,416c | le Sage et al.; Schmidt et al.; Li et al.; Hsu et al.; Thege et al.; Yang et al.; Deng et al.; Wang et al.28,31,66,67,68,69,70,71 | ||
| VPR (MS2-p65-HSF1) (SAM) | ∼5,799 + ∼1,416c | Giehrl-Schwab et al.72 | ||
| VPH | ∼5,745 | Tian et al.27 | ||
| VPH-dSpCas9-SS18 | ∼6,471 | Tian et al.; Beyersdorf et al.27,73 | ||
| dxCas9 (dSpCas9 derivative) | 5′-NG-3′, 5′-GAA-3′, 5′-GAT-3′ |
VPR | ∼5,799 | Hu et al.74 |
| dSaCas9 | 5′-NNGRRT-3′ | – | ∼3,159 | |
| KRAB | ∼3,399 | Escobar et al.; Black et al.; Gemberling et al.38,45,75 | ||
| VP64 | ∼3,438 | Matharu et al.44 | ||
| VPR | ∼4,854 | Jensen et al.; Cui et al.; Escobar et al.; Nguyen et al.; Di Maria et al.26,32,38,54,58 | ||
| VPR (Mini) | ∼4,104 | Vora et al.76 | ||
| dAsCas12a | 5′-TTTV-3′ | – | ∼3,921 | |
| KRAB | ∼4,314 | Escobar et al.38 | ||
| VPR | ∼5,658 | Escobar et al.38 | ||
| Sp65p3-HSF1 (Activ) | ∼4,659 | Campa et al.77 | ||
| dLbCas12a | 5′-TTTV-3′ | – | ∼3,684 | |
| KRAB | ∼4,077 | Escobar et al.38 | ||
| VPR | ∼5,421 | Escobar et al.38 | ||
| dimpLbCas12a (dLbCas12a-RVRR) | 5′-TNTN-3′ 5′-TACV-3′ 5′-TTCV-3′ 5′-CTCV-3′ 5′-CCCV-3′ |
SunTag (GCN4-VP64) | ∼4,644 + ∼2,034b | Tóth et al.78 |
| enAsCas12a | 5′-TTYN-3′ 5′-VTTV-3′ 5′-TRTV-3′ |
VPR | 5,658 | Kleinstiver et al.25 |
a
Many variations of VP16 repeats exist and have been used for CRISPRa, such as VP48, VP160, and VP192, but the most utilized is the VP64 variant.
b
The first number indicates the size of the dCas-Suntag construct with 10xGCN4 binding sites, and the second denotes the size of the scFv-containing construct.
c
The first number is the size of the dSpCas9-effector construct, the second is the size of the MS2-p65-HSF1 construct.