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

A subset of these plasmids is included in the CRISPRecise Kit.

Plasmids from Article

ID	Plasmid	Purpose
126753	pX330-Flag-WT_SpCas9 (without sgRNA; with silent mutations)	Expression plasmid for human codon-optimized WT SpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126754	pX330-Flag-eSpCas9 (without sgRNA; with silent mutations)	Expression plasmid for human codon-optimized increased fidelity eSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126755	pX330-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)
126756	pX330-Flag-HypaSpCas9 (without sgRNA; with silent mutations)	Expression plasmid for human codon-optimized increased fidelity HypaSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126758	pX330-Flag-evoSpCas9 (without sgRNA; with silent mutations)	Expression plasmid for human codon-optimized increased fidelity evoSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126759	pX330-Flag-HeFSpCas9 (without sgRNA; with silent mutations)	Expression plasmid for human codon-optimized increased fidelity HeFSpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126760	B-SpCas9	Expression 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.
126761	B-eSpCas9	Expression 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.
126762	B-SpCas9-HF1	Expression 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.
126763	B-HypaSpCas9	Expression 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.
126765	B-evoSpCas9	Expression 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.
126766	B-HeFSpCas9	Expression 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.
126767	eSpCas9-plus	Expression 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.
126768	SpCas9-HF1-plus	Expression 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
126769	pET-FLAG-eSpCas9	Expression of increased fidelity eSpCas9 in bacterial cells
126770	pET-FLAG-SpCas9-HF1	Expression of increased fidelity SpCas9-HF1 in bacterial cells
126772	pET-FLAG-B-eSpCas9	Expression 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
126774	pET-FLAG-eSpCas9-plus	Expression 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.
126775	pET-FLAG-SpCas9-HF1-plus	Expression 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.
126776	pmCherry_sgRNA-ver2	gRNA expression vector with mCherry transfection control, does not contain the direct repeat of the truncated guideRNA
126777	pX330-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)
126778	pX330-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)