Table 1.
Main studies that have used dCas9-based CRISPRa/i systems for gene activation or repression.
| CRISPRa/i system | Target gene (s) | In vitro/In vivo | Outcome | Reference |
|---|---|---|---|---|
| dCas9-VP64 | VEGFA, NTF3 (human) | In vitro (human HEK293) | Activation | (Maeder et al., 2013) |
| dCas9-VP64 | IL1RN, NANOG, HBG1/2, MYOD1, VEGFA, TERT, IL1B, IL1R2 (human) ASCL1 (human and Mus musculus) |
In vitro (human HEK293T, mouse primary embryonic fibroblasts) | Activation | (Perez-Pinera et al., 2013) |
| dCas9-VP64 | SIM1, MC4R (Mus musculus) | In vitro (mouse Neuro-2A) and in vivo (mouse) | Activation | (Matharu et al., 2019) |
| dCas9-VPR | MIAT, NEUROD1, ASCL1, RHOXF2, TTN, ACTC1, NGN2 (human) ACTA1, ACTC1, TTN, TUNA (Mus musculus) MTK, CECA1 (D. melanogaster) GAL7, HED1 (S. cerevisae) |
In vitro (human HEK293T, mouse Neuro-2A, Drosophila S2R+, yeast strain W303) | Activation | (Chavez et al., 2015) |
| dCas9-VPR | MEF2D, KLF15 (Mus musculus) | In vivo (mouse heart) | Activation | (Schoger et al., 2020) |
| dCas9-SAM | ASCL1, MYOD1, NEUROG2, VEGFA, HBG1, TERT, IL1B, IL1R2, MYC, ZFP42, LIN28A, SOX2, NANOG, KLF4, POU5F1, long intergenic non-coding RNAs (lincRNA) (human) | In vitro (human HEK293FT, A375) | Activation | (Konermann et al., 2015) |
| dCas9-SunTag-VP64 | CXCR4, CDKN1B (human) | In vitro (human HEK293, U2OS, K562) | Activation | (Tanenbaum et al., 2014) |
| dCas9-SunTag-VP64 | Reactivation of latent HIV-1 (human) | In vitro (human C11, J-Lat, ACH2, HEK293T, TZM-bl) | Activation | (Ji et al., 2016) |
| dCas9-SunTag-VP64 | MYC, TNFRSF1A, SLC7A11, TP53 (Mus musculus) | In vivo (mouse liver) | Activation | (Wangensteen et al., 2018) |
| dCas9-SunTag-TET1CD | STAT3-binding site of GFAP, H19, RHOXF2B, CARD9, SH3BP2, CNKSR1 (Mus musculus) | In vitro (mouse embryonic stem cells, neuronal precursor cells, A549) and in vivo (mouse brain) | Activation | (Morita et al., 2016) |
| dCas9-SunTag-p65-HSF1 | Exogenous mCherry, endogenous genes ASCL1, NEUROG2, NEUROD1, ACTA1, DKK1, SLC6A4, RNF43, BCL2, ZNRF3, PRDM16, HBB, GRM2, LNS2, IL10, SLC7A11 and lncRNAs MIAT, HALGR, FENDRR, LNCPINT (Mus musculus) | In vitro (human HEK293T, mouse Neuro-2A, primary astrocytes and embryonic fibroblasts) and in vivo (mouse brain) | Activation | (Zhou et al., 2018) |
| dCas9-VPR, SAM, SunTag-VP64 | TTN, HBG1, MIAT, TUNAR, RHOXF2, ACTC1, ASCL1, NEUROD1, CXCR4 (human) HBB-BH1, TTN (Mus musculus) Wingless, Twist (D. melanogaster) |
In vitro (human HEK293T, HeLa, U2OS and MCF7, mouse Neuro-2A and NIH-3T3, Drosophila S2R+) | Activation | (Chavez et al., 2016) |
| dCas9-VP64, dCas9-p300 | IL1RN, MYOD, OCT4, mammalian beta-globin locus control region (HBE, HBG, HBD, HBB) (human) | In vitro (human HEK293T) | Activation | (Hilton et al., 2015) |
| dCas9-TET1CD | BRCA1 (human) | In vitro (human HeLa, MCF7) | Activation | (Choudhury et al., 2016) |
| High-fidelity dCas9-TET3CD | RASAL1, EYA1, LRFN2, KLOTHO (human) | In vitro (human kidney cells, TK173, TK188 fibroblasts, and HK2 epithelial cells, mouse primary kidney fibroblasts and renal tubular epithelial cells) and in vivo (mouse kidney) | Activation | (Xu et al., 2018) |
| dCas9-KRAB | CD71, CXCR4 (human) | In vitro (human HEK293, HeLa) | Repression | (Gilbert et al., 2013) |
| dCas9-KRAB | CALM1, 2, 3 (human) | In vitro (human iPSC-CMs) | Repression | (Limpitikul et al., 2017) |
| dCas9-KRAB | Including GAS5, H19, MALAT1, NEAT1, TERC, XIST (lncRNAs) (human) | In vitro (human K562) | Repression | (Gilbert et al., 2014) |
| dCas9-KRAB | SYT1, VAMP2, SNAP25, STX1A, STX1B, DOC2A, DOC2B (Mus musculus) | In vitro (mouse primary neurons) and in vivo (mouse brain) | Repression | (Zheng et al., 2018) |
| dCas9-KRAB, dCas9-KRAB-MeCP2 | CANX, CXCR4, CHK1, SEL1L, ARPC2, MAPK3, BRCA1, SYVN1, BLM, GZMM, MAPK3, RHOA, CHEK1, CHEK2, ARPC2, TERC, XIST (human) | In vitro (human HEK293T, HAP1, SH-SY5Y) | Repression | (Yeo et al., 2018) |
| dCas9-LSD1, dCas9-KRAB | OCT4, TBX3 (Mus musculus) | In vitro (mouse embryonic stem cells) | Repression | (Kearns et al., 2015) |
| dCas9-SunTag-DNMT3A | HOX5A, BACH2, KLF4 (human) | In vitro (human HEK293T) | Repression | (Huang et al., 2017) |
| dCas9-DNMT3A, dCas9-SunTag-DNMT3A | UNC5C, CCDC85C intron, SHB intron, MIR152, GAD1 intron 3, NRF1 binding sites (human) | In vitro (human HEK293T, HeLa, MCF7) | Repression | (Pflueger et al., 2018) |
| dCas9-DNMT3A-DNMT3L (DNMT3A3L) | EpCAM, CXCR4, TFRC (human) | In vitro (human SKOV-3, HEK293) | Repression | (Stepper et al., 2017) |
| VP64-dCas9-VP64 | BRN2, ASCL1,MYT1L (Mus musculus) | In vitro (mouse embryonic fibroblasts) | Reprogramming mouse embryonic fibroblasts into induced neuronal cells | (Black et al., 2016) |
| dCas9-TET1, dCas9-DNMT3A | BDNF, MYOD (Mus musculus) | In vitro (mouse C3H10T1/2, embryonic stem cells, fibroblasts) and in vivo (mouse brain, skin epidermis) | Reprogramming fibroblasts into myoblasts | (Liu et al., 2016) |
| dCas9-SunTag-VP64/p300 | OCT4, SOX2, KLF4, c-MYC, NR5A2, GLIS1, CEBPA (Mus musculus) | In vitro (mouse tail tip fibroblasts, embryonic fibroblasts) | Reprogramming mouse embryonic fibroblasts into iPSCs | (Liu et al., 2018) |
| dCas9-SAM | GATA4, HAND2, MEF2C TBX5 (human) | In vitro (human foreskin fibroblasts) | Reprogramming human fibroblasts into induced cardiac progenitor cells (iCPCs) | (Wang et al., 2020) |
| dCas9-SAM | GATA4, NKX2–5, TBX5, HAND2, MESP1, BAF60C, ISL1, GATA6, SRF, HAND1/2, IRX4 (Mus musculus) | In vitro (mouse primary fibroblasts) | Reprogramming mouse tail-tip fibroblasts into CRISPR-induced cardiovascular progenitor cells (ciCPCs) | (Jiang et al., 2022) |
| dCas9-VP64 | GATA4, NKX2–5, MEF2C, HAND2 and TNNT2 (Rattus norvegicus) | In vitro (primary neonatal rat hearts to generate cardiospheres) and in vivo (rat heart) | Cardio-specific differentiation factor activation in cardiosphere-derived cells (CDCs) | (Sano et al., 2022) |