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Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways.
Taghbalout A, Du M, Jillette N, Rosikiewicz W, Rath A, Heinen CD, Li S, Cheng AW
Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. PubMed Article

Plasmids from Article

ID Plasmid Purpose
134632pAT243_pX-sgRNA-5xPBSa-5xPBScCloning vector for expression of sgRNAs with dual PUF-Binding Sites (5xPBSa - 5xPBSc) of Casilio-ME 2.3, 2.4, 3.3 and 3.4
134643pAT888_CMV/CAG_PUFa-hTET1(CD)3rd generation lentiviral expression of untagged Casilio-ME1 effector PUFa-hTET1(CD) in mammalian cells
134649pAT890_CMV/CAG_dCas9-hTET1(CD)3rd generation lentiviral plasmid for expression of Sp dCas9-hTET1(CD) fusion protein in mammalian cells
134650pAT801_CMV/CAG_TALE_A-hTET1(CD)3rd generation lentiviral expression of TALE-hTET1(CD) in mammalian cells targeting MLH1 promoter region
134740pAT812_CMV/CAG_TALE_B-hTET1(CD)3rd generation lentiviral expression of TALE-hTET1(CD) in mammalian cells targeting MLH1 promoter region
134829pAT806_CMV/CAG_TALE_D-hTET1(CD)3rd generation lentiviral expression of TALE-hTET1(CD) in mammalian cells targeting MLH1 promoter region
134830pAT817_CMV/CAG_TALE_F-hTET1(CD)3rd generation lentiviral expression of TALE-hTET1(CD) in mammalian cells targeting MLH1 promoter region
134831pAT341_CMV/CAG_PUFa-TET1CD (H1671Y D1673A) (dead TET1)3rd generation lentiviral expression of catalytically inactive Casilio-ME1 effector containing dead hTET1(CD) H1671Y D1673A in mammalian cells
134835pAT360_CMV/CAG_PUFa-hGADD45A-hTET1(CD)3rd generation lentiviral expression of untagged Casilio-ME2.1 effector PUFa-hGADD45A-hTET1(CD) in mammalian cells
134837pAT635_CMV/CAG_hGADD45A-PUFa-hTET1(CD)3rd generation lentiviral expression of untagged Casilio-ME2.2 effector hGADD45A-PUFa-hTET1(CD) in mammalian cells
134838pAT355_CMV/CAG_PUFc-hGADD45A3rd generation lentiviral expression of Casilio-ME2.3 effector PUFc-hGADD45A in mammalian cells
134846pAT356-CMV/CAG_hGADD45A-PUFc3rd generation lentiviral expression of Casilio-ME2.4 effector hGADD45A-PUFc in mammalian cells
134848pAT892_CMV/CAG_PUFa-p65HSF13rd generation lentiviral expression of Casilio activator PUFa-p65HSF1 in mammalian cells
134849pAT608_CMV/CAG_Flag-hNEIL2-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector hNEIL2-PUFa-hTET1(CD) in mammalian cells
134850pAT611_CMV/CAG_PUFa-hNEIL2-hTET1(CD)3rd generation lentiviral expression of Casilio-ME3.2 effector PUFa-hNEIL2-hTET1(CD) in mammalian cells
134851pAT595_CMV/CAG_PUFc-hNEIL23rd generation lentiviral expression of Casilio-ME3.3 effector PUFc-hNEIL2 in mammalian cells
134855pAT596_CMV/CAG_hNEIL2-PUFc3rd generation lentiviral expression of Casilio-ME3.4 effector hNEIL2-PUFc in mammalian cells
134856pAT699-CMV/CAG_Flag-hGADD45A-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector hGADD45A-PUFa-hTET1(CD) in mammalian cells
134870pAT976_CMV/CAG_Flag-hGADD45A-PUFa-dTET1CD3rd generation lentiviral expression of Flag-tagged and catalytically inactive Casilio-ME2.2 effector containing dead hTET1(CD) H1671Y D1673A in mammalian cells
134871pAT971_CMV/CAG_Flag-hGADD45A (G39A)-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with G39A GADD45A mutation hGADD45A(G39A)-PUFa-hTET1(CD) in mammalian cells
134872pAT972_CMV/CAG_Flag-hGADD45A (L77E)-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with L77E GADD45A mutation hGADD45A (L77E)-PUFa-hTET1(CD) in mammalian cells
134873pAT973_CMV/CAG_FlaghGADD45A (G39A, L77E)-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with G39A L77E GADD45A mutations hGADD45A(G39A L77E)-PUFa-hTET1(CD) in mammalian cells
134875pAT977_CMV/CAG_Flag-hNEIL2-PUFa-dTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector containing a catalytically dead hTET1(CD) in mammalian cells
134876pAT1059_CMV/CAG_Flag-hNEIL2 (C291S)-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector with C291S NEIL2 mutation hNEIL2(C291S)-PUFa-hTET1(CD) in mammalian cells
134877pAT1060_CMV/CAG_Flag-hNEIL2 (R310Q)-PUFa-hTET1(CD)3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector with R310Q NEIL2 mutation hNEIL2(R310-Q)-PUFa-hTET1(CD) in mammalian cells
134895pAT1089_PB-EF1a-Blast_2A_rtTA3-SV40pA_pCW-dCas9PiggyBac vector for establishement of doxycycline-inducible Sp dCas9 expression cell lines
134975pAT1501-[PB-4xHSab-EF1a-Hygro_2A_rtTA3-SV40pA(-)_pCW-DEST]Destination vector for doxycycline-inducible expression from plasmid hosting piggyBac hygromycin cassettes
134976pAT1502-[PB-4xHSab-EF1a-Blast_2A_rtTA3-SV40pA(-)_pCW-DEST]Destination vector for doxycycline-inducible expression from plasmid hosting piggyBac Blasticidin cassettes
134978pAT1503_pX-SPFE-BbsI(ccdbCam)-5xPBSaCloning vector for expression of Casilio sgRNAs containing five PUFa-binding sites (PBSa)
134979pAT1504_pX-SPFE-BbsI(ccdbCam)-5xPBScCloning vector for expression of Casilio sgRNAs containing five PUFc-binding sites (PBSc)

Antibodies from Article