From: DNA methylation and de-methylation using hybrid site-targeting proteins
Strategy | Delivery | Efficacy | Duration | Locus | Peak site | Editing range | Target | Off-targets | Features | Refs |
---|---|---|---|---|---|---|---|---|---|---|
TALE-DNMT3ACD-3 L | Transfection Transduction | 12–15% 17% | 2 days 4 days | CDKN2A | n.a. | ~ 700 bp | CpG | At nearby regions (Targeted mePCR) | First TALE-DNMT3 A study (2015) | [33] |
TALE-CIB1; DNMT3A/TET1 CD-C R Y2 | Transfection | ~ 10% | ~ 5 days | Ascl 1 | n.a. | n.a. | CpG | n.a. | Epigenome editing by optogenetics (2017) | [34] |
dCas9-DNMT3ACD | Transfection | ~ 35% ~ 43% | 8 days 3 days | BACH2 CDKN2A | −27 bp, + 25 bp | < 100 bp | CpG | Off-targets observed using unspeci_c sgRNAs | First dCas9-DNMT3 A studies (2016) | |
dCas9-DNMT3A | Transduction | ~ 12% ~ 25% | 3 days 3 days | Gapdhsnrpn CDKN2A | n.a. | n.a. | CpG | Not detectable (dCas9 ChIP-seq) | Targeted methylation alters CTCF looping and local genes expression (2016) | [40] |
TALE/dCas9-KRAB, DNMT3ACD, DNMT3L | Transduction | ~ 80% | 30 days | B2M | n.a. | ~ 3 kb | CpG | Not detectable (MeDIP-seq) | Inheritable silencing of endogenous genes (2016) | [43] |
dCas9-DNMT3ACD-3 L | Transfection | ~ 30% | 5 days | TFRC CXCR4 | −40 bp, + 25 bp | ~ 800 bp | CpG | Mild off-target observed | More e_cient than dCas9- DNMT3ACD (2016) | [44] |
dCas9-Sun Tag-DNMT3A | Transduction | ~ 70% ~ 45% | 30 days | HOXA5 DLX1 | n.a. | ~ 4.5 kb ~ 500 bp | CpG (CpH) | Not detectable (RRBS; WGBS) | Long range methylation Non-CpG methylation (2017) | [47] |
dCas9-Sun Tag-DNMT3ACD | Transfection | ~ 15% | 4 days | UNC5C | n.a. | n.a. | CpG | Not detectable (TSC-bs-seq) | Modular Sun Tag- DNMT3ACD reduces off-target events (2018) | [62] |
dCas9-MQ1 147 | Transfection | *60% | 24 h | HOXA5 HOXA4 RUNX1 | −24-26 bp | n.a. | CpG | Not detectable (RRBS) | In vivo application using zygote injection (2017) | [54] |
dCas9-split M.SssI | Transfection | *50% | 2 days | SALL2 | −22-23 bp | 8–25 bp | CpG | Not detectable (Targeted mePCR) | Split catalytic domain for higher speci_cty (2017) | [48] |