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A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets.

Kulcsar PI, Talas A, Ligeti Z, Toth E, Rakvacs Z, Bartos Z, Krausz SL, Welker A, Vegi VL, Huszar K, Welker E
Nat Commun. 2023 Sep 16;14(1):5746. doi: 10.1038/s41467-023-41393-5. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
126757HypaR-SpCas9Expression plasmid for human codon-optimized increased fidelity Hypa2SpCas9 (without U6-sgRNA coding sequence, with silent mutations)
126764B-HypaR-SpCas9Expression plasmid for human codon-opt. increased fidelity Blackjack-Hypa2SpCas9 (w/o U6-sgRNA). Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than Hypa2SpCas9.
207361B-Sniper SpCas9Expression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207362B-HiFi SpCas9Expression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207363B-evoSpCas9-V495MExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207364B-evoSpCas9-N515YExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207365B-evoSpCas9-E526KExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207366B-evoSpCas9-Q661RExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207367B-HeFSpCas9-A661RExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207368B-HeFSpCas9-A695QExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207369B-HeFSpCas9-A848KExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207370B-HeFSpCas9-A926QExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207371B-HeFSpCas9-A1003KExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207372B-HeFSpCas9-A1060RExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
207373PET-WT SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207374PET-Sniper SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207375PET-Blackjack SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207376PET-HiFi SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207377PET-B-Sniper SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207378PET-B-HiFi SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207379PET-eSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207380PET-eSpCas9-plus-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207381PET-SpCas9-HF1-plus-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207382PET-SpCas9-HF1-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207383PET-B-eSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207384PET-HypaSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207385PET-B-SpCas9-HF1-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207386PET-B-HypaSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207387PET-HypaR-SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207388PET-B-HypaR-SpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207389PET-evoSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207390PET-B-evoSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207391PET-HeFSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells
207392PET-B-HeFSpCas9-NLS-6xHisExpression of increased-fidelity (i.e. high-fidelity) SpCas9 variant in bacterial cells

Antibodies from Article