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Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5'G-extended sgRNAs.
Kulcsar PI, Talas A, Toth E, Nyeste A, Ligeti Z, Welker Z, Welker E
Nat Commun. 2020 Mar 6;11(1):1223. doi: 10.1038/s41467-020-15021-5.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
126753pX330-Flag-WT_SpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized WT SpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126754pX330-Flag-eSpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity eSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126755pX330-Flag-SpCas9-HF1 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity SpCas9-HF1 (without U6-sgRNA coding sequence, with silent mutations)
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126756pX330-Flag-HypaSpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity HypaSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126758pX330-Flag-evoSpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity evoSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126759pX330-Flag-HeFSpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity HeFSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126760B-SpCas9Expression plasmid for human codon-optimized Blackjack-SpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than that of WT SpCas9.
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126761B-eSpCas9Expression plasmid for human codon-optimized increased fidelity Blackjack-eSpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than eSpCas9.
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126762B-SpCas9-HF1Expression plasmid for human codon-opt. increased fidelity Blackjack-SpCas9-HF1 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than SpCas9HF1.
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126763B-HypaSpCas9Expression plasmid for human codon-opt. increased fidelity Blackjack-HypaSpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than HypaSpCas9.
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126765B-evoSpCas9Expression plasmid for human codon-opt. increased fidelity Blackjack-evoSpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than evoSpCas9.
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126766B-HeFSpCas9Expression plasmid for human codon-opt. increased fidelity Blackjack-HeFSpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than HeFSpCas9.
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126767eSpCas9-plusExpression plasmid for human codon-optimized increased fidelity eSpCas9-plus (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as eSpCas9.
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126768SpCas9-HF1-plusExpression plasmid for human codon-opt. increased fidelity SpCas9-HF1-plus (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as SpCas9-HF1
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126769pET-FLAG-eSpCas9Expression of increased fidelity eSpCas9 in bacterial cells
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126770pET-FLAG-SpCas9-HF1Expression of increased fidelity SpCas9-HF1 in bacterial cells
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126772pET-FLAG-B-eSpCas9Expression of increased fidelity Blackjack-eSpCas9 in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than that of eSpCas9
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126774pET-FLAG-eSpCas9-plusExpression of increased fidelity eSpCas9-plus in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as eSpCas9.
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126775pET-FLAG-SpCas9-HF1-plusExpression of increased fidelity SpCas9-HF1-plus in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as SpCas9-HF1.
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126776pmCherry_sgRNA-ver2gRNA expression vector with mCherry transfection control, does not contain the direct repeat of the truncated guideRNA
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126777pX330-Flag-Sniper SpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity Sniper SpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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126778pX330-Flag-HiFi SpCas9 (without sgRNA; with silent mutations)Expression plasmid for human codon-optimized increased fidelity HiFi SpCas9 (without U6-sgRNA coding sequence, with silent mutations)
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