Engineered CRISPR-OsCas12f1 and RhCas12f1 with robust activities and expanded target range for genome editing.

Kong X, Zhang H, Li G, Wang Z, Kong X, Wang L, Xue M, Zhang W, Wang Y, Lin J, Zhou J, Shen X, Wei Y, Zhong N, Bai W, Yuan Y, Shi L, Zhou Y, Yang H
Nat Commun. 2023 Apr 11;14(1):2046. doi: 10.1038/s41467-023-37829-7. PubMed Article

Plasmids from Article

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ID	Plasmid	Purpose
197031	pCBh_3x Flag-NLS-denOsCas12f1-NLS-VPR-NLS-mCherry_pA_phU6_gRNA scaffold-spacer_pCMV_BFP_pA	Vector encoding a human codon-optimized dead enOsCas12f1 (denOsCas12f1)-VPR driven by CBh promoter, guide RNAs compatible with enOsCas12f1 driven by hU6, and BFP driven by CMV promoter
197030	miniCRISPRoff-v1	Vector encoding a human codon-optimized miniCRISPRoff-v1 driven by CBh promoter, guide RNAs compatible with enOsCas12f1 driven by hU6, and mCherry driven by CMV promoter
197029	AAV2 ITR_phU6_gRNA scaffold-DMD sg1_phU6_gRNA scaffold-DMD sg16_pMHCK7-3x Flag-NLS-ecDHFR-enOsCas12f1-NLS-ecDHFR_pA_AAV2 ITR	AAV vector encoding a human codon-optimized ecDHFR-enOsCas12f1-ecDHFR driven by MHCK7 promoter, DMD-targeting guide RNAs compatible with enOsCas12f1 driven by hU6
197028	AAV2 ITR_phU6_gRNA scaffold-DMD sg1_phU6_gRNA scaffold-DMD sg16_pMHCK7-3x Flag-NLS-enOsCas12f1-NLS_pA_AAV2 ITR	AAV vector encoding a human codon-optimized enOsCas12f1 driven by MHCK7 promoter, DMD-targeting guide RNAs compatible with enOsCas12f1 driven by hU6
197027	pCBh_3x Flag-NLS-enRhCas12f1-NLS_pA_phU6_gRNA scaffold-spacer_pCMV_mCherry_pA	Vector encoding a human codon-optimized enRhCas12f1 driven by CBh promoter, guide RNAs compatible with enRhCas12f1 driven by hU6, and mCherry driven by CMV promoter
197026	pCBh_3x Flag-NLS-enOsCas12f1-NLS_pA_phU6_gRNA scaffold-spacer_pCMV_mCherry_pA	Vector encoding a human codon-optimized enOsCas12f1 driven by CBh promoter, guide RNAs compatible with enOsCas12f1 driven by hU6, and mCherry driven by CMV promoter
197025	pCBh_3x Flag-NLS-RhCas12f1-NLS_pA_phU6_gRNA scaffold-spacer_pCMV_mCherry_pA	Vector encoding a human codon-optimized RhCas12f1 driven by CBh promoter, guide RNAs compatible with RhCas12f1 driven by hU6, and mCherry driven by CMV promoter
197024	pCBh_3x Flag-NLS-OsCas12f1-NLS_pA_phU6_gRNA scaffold-spacer_pCMV_mCherry_pA	Vector encoding a human codon-optimized OsCas12f1 driven by CBh promoter, guide RNAs compatible with OsCas12f1 driven by hU6, and mCherry driven by CMV promoter