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Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing.

Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, Aryee MJ, Joung JK
Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 10.1038/s41587-018-0011-0. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
107941pCAG-enAsCas12a(E174R/S542R/K548R)-NLS(nuc)-3xHA (AAS848)Mammalian expression plasmid for human codon optimized enAsCas12a (enhanced AsCas12a) encoding E174R/S542R/K548R substitutions
107942pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815)Mammalian expression plasmid for human codon optimized enAsCas12a-HF1 (enhanced high-fidelity AsCas12a) encoding E174R/N282A/S542R/K548R substitutions
107943pCAG-denAsCas12a(E174R/S542R/K548R/D908A)-NLS(nuc)-3xHA-VPR (RTW776)Mammalian expression plasmid for human codon optimized DNase-inactive enAsCas12a (enhanced AsCas12a) fused to the VPR activation domain, encoding E174R/S542R/K548R/D908A substitutions
114067pT7-G-AsCas12a crRNA-BsaI cassette (MSP3491)T7 promoter crRNA entry vector used for in vitro transcription of AsCas12a guides
114068pT7-G-LbCas12a crRNA-BsaI cassette (MSP3495)T7 promoter crRNA entry vector used for in vitro transcription of LbCas12a guides
114069pET-28b-T7-hAsCas12a-NLS(nucleoplasmin)-6xHis (BPK3541)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a with C-terminal NLS and His-tag
114070pET-28b-T7-bLbCas12a-NLS(nucleoplasmin)-6xHis (RTW645)T7 promoter bacterial expression plasmid for bacterial codon optimized LbCas12a with C-terminal NLS and His-tag
114071pET-28b-T7-hAsCas12a(E174R/S542R)-NLS(nucleoplasmin)-6xHis (AAS1880)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a(E174R/S542R) with C-terminal NLS and His-tag
114072pET-28b-T7-henAsCas12a(E174R/S542R/K548R)-NLS(nucleoplasmin)-6xHis (AAS1885)T7 promoter bacterial expression plasmid for human codon optimized enAsCas12a(E174R/S542R/K548R) with C-terminal NLS and His-tag
114073pET-28b-T7-henAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nucleoplasmin)-6xHis (AAS1935)T7 promoter bacterial expression plasmid for human codon optimized enAsCas12a-HF1(E174R/N282A/S542R/K548R) with C-terminal NLS and His-tag
114074pET-28b-T7-hAsCas12a-RVR(S542R/K548V/N552R)-NLS(nucleoplasmin)-6xHis (AAS1897)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a-RVR(S542R/K548V/N552R) with C-terminal NLS and His-tag
114075pET-28b-T7-hAsCas12a-enRVR(E174R/S542R/K548V/N552R)-NLS(nucleoplasmin)-6xHis (AAS1931)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a-enRVR(E174R/S542R/K548V/N552R) with C-terminal NLS and His-tag
114076pET-28b-T7-hAsCas12a-RR(S542R/K607R)-NLS(nucleoplasmin)-6xHis (AAS1916)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a-RR(S542R/K607R) with C-terminal NLS and His-tag
114077pET-28b-T7-hAsCas12a-enRR(E174R/S542R/K607R)-NLS(nucleoplasmin)-6xHis (AAS1902)T7 promoter bacterial expression plasmid for human codon optimized AsCas12a-enRR(E174R/S542R/K607R) with C-terminal NLS and His-tag
114078pCAG-hdAsCas12a(D908A)-NLS(nucleoplasmin)-gs-3xHA-gs-VPR (RTW876)CAG promoter expression plasmid for human codon optimized AsCas12a-VPR(1.1)
114079pCAG-rAPOBEC1-gs-XTEN-gs-hdAsCas12a(D908A)-NLS(nucleoplasmin)-gs-UGI-NLS(SV40)(AsBE1.1) (RTW1351)CAG promoter expression plasmid for human codon optimized AsBE1.1
114080pCAG-NLS(SV40)x2-rAPOBEC1-gs-XTEN-gs-hdAsCas12a(D908A)-gs-UGI-NLS(SV40)(AsBE1.4) (RTW1008)CAG promoter expression plasmid for human codon optimized AsBE1.4
114081pCAG-rAPOBEC1-gs-XTEN-gs-hdenAsCas12a(E174R/S542R/K548R/D908A)-NLS(nucleoplasmin)-gs-UGI-NLS(SV40)(enAsBE1.1) (RTW1352)CAG promoter expression plasmid for human codon optimized enAsBE1.1
114082pCAG-NLS(SV40)x2-rAPOBEC1-gs-XTEN-gs-hdenAsCas12a(E174R/S542R/K548R/D908A)-NLS(nucleoplasmin)-gs-UGI-NLS(SV40)(enAsBE1.2)CAG promoter expression plasmid for human codon optimized enAsBE1.2, also called RTW1348
114083pCAG-rAPOBEC1-gs-XTEN-gs-hdenAsCas12a(E174R/S542R/K548R/D908A)-gs-UGI-NLS(SV40)(enAsBE1.3) (RTW1296)CAG promoter expression plasmid for human codon optimized enAsBE1.3
114084pCAG-NLS(SV40)x2-rAPOBEC1-gs-XTEN-gs-hdenAsCas12a(E174R/S542R/K548R/D908A)-gs-UGI-NLS(SV40)(enAsBE1.4) (RTW1028)CAG promoter expression plasmid for human codon optimized enAsBE1.4
114085pCAG-rAPOBEC1-gs-XTEN-gs-hdLbCas12a(D832A)-NLS(nucleoplasmin)-gs-UGI-NLS(SV40)(LbBE1.1) (RTW1295)CAG promoter expression plasmid for human codon optimized LbBE1.1
114086pCAG-NLS(SV40)x2-rAPOBEC1-gs-XTEN-gs-hdLbCas12a(D832A)-gs-UGI-NLS(SV40)(LbBE1.4) (RTW1293)CAG promoter expression plasmid for human codon optimized LbBE1.4
114087pUC19-U6-FnCas12a crRNA-BsmBI cassette (BPK4446)U6 promoter crRNA entry vector used for all FnCas12a crRNAs (clone spacer oligos into BsmBI cassette)
114088pUC19-U6-MbCas12a crRNA-BsmBI cassette (BPK4449)U6 promoter crRNA entry vector used for all MbCas12a crRNAs (clone spacer oligos into BsmBI cassette)
114089pCAG-hFnCas12a-NLS(nucleoplasmin)-3xHA (AAS1472)CAG promoter expression plasmid for human codon optimized FnCas12a nuclease with C-terminal NLS and HA tag
114090pCAG-hMbCas12a-NLS(nucleoplasmin)-3xHA (AAS2134)CAG promoter expression plasmid for human codon optimized MbCas12a nuclease with C-terminal NLS and HA tag
114091pCAG-hAsCas12a(E174R/S542R)-NLS(nucleoplasmin)-3xHA (AAS826)CAG promoter expression plasmid for human codon optimized AsCas12a(E174R/S542R) nuclease with C-terminal NLS and HA tag
114092pCAG-hAsCas12a-RVR(S542R/K548V/N552R)-NLS(nucleoplasmin)-3xHA (AAS1065)CAG promoter expression plasmid for human codon optimized AsCas12a-RVR(S542R/K548V/N552R) nuclease with C-terminal NLS and HA tag
114093pCAG-hAsCas12a-enRVR(E174R/S542R/K548V/N552R)-NLS(nucleoplasmin)-3xHA (AAS1060)CAG promoter expression plasmid for human codon optimized AsCas12a-enRVR(E174R/S542R/K548V/N552R) nuclease with C-terminal NLS and HA tag
114094pCAG-hAsCas12a-RR(S542R/K607R)-NLS(nucleoplasmin)-3xHA (AAS1081)CAG promoter expression plasmid for human codon optimized AsCas12a-RR(S542R/K607R) nuclease with C-terminal NLS and HA tag
114095pCAG-hAsCas12a-enRR(E174R/S542R/K607R)-NLS(nucleoplasmin)-3xHA (AAS1077)CAG promoter expression plasmid for human codon optimized AsCas12a-enRR(E174R/S542R/K607R) nuclease with C-terminal NLS and HA tag

Antibodies from Article