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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.
PubMed 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
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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
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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
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114067pT7-G-AsCas12a crRNA-BsaI cassette (MSP3491)T7 promoter crRNA entry vector used for in vitro transcription of AsCas12a guides
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114068pT7-G-LbCas12a crRNA-BsaI cassette (MSP3495)T7 promoter crRNA entry vector used for in vitro transcription of LbCas12a guides
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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
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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
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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
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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
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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
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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
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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
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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
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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
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114078pCAG-hdAsCas12a(D908A)-NLS(nucleoplasmin)-gs-3xHA-gs-VPR (RTW876)CAG promoter expression plasmid for human codon optimized AsCas12a-VPR(1.1)
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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
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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
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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
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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
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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
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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
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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
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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
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114087pUC19-U6-FnCas12a crRNA-BsmBI cassette (BPK4446)U6 promoter crRNA entry vector used for all FnCas12a crRNAs (clone spacer oligos into BsmBI cassette)
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114088pUC19-U6-MbCas12a crRNA-BsmBI cassette (BPK4449)U6 promoter crRNA entry vector used for all MbCas12a crRNAs (clone spacer oligos into BsmBI cassette)
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114089pCAG-hFnCas12a-NLS(nucleoplasmin)-3xHA (AAS1472)CAG promoter expression plasmid for human codon optimized FnCas12a nuclease with C-terminal NLS and HA tag
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114090pCAG-hMbCas12a-NLS(nucleoplasmin)-3xHA (AAS2134)CAG promoter expression plasmid for human codon optimized MbCas12a nuclease with C-terminal NLS and HA tag
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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
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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
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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
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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
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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
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