| 姓名 | 描述 | Application Notes | PAM Sequence | References | 序列 |
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| zCas9 | Zebrafish codon-optimized version of S. pyogenes Cas9 with SV40 large T-antigen nuclear localization signals (nls) at both its amino and carboxyl termini | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci USA. 110: 13904 (2013) |
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| hCas9 | Human codon-optimized CRISPR associated protein 9 from Streptococcus pyogenes (with codon-optimized 3xFLAG tag and nuclear localization signal), also known as SpCas9 | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Science. 339:819 (2013) |
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| Cas9(D10A) | Human codon-optimized CRISPR associated protein 9 with D10A mutation, HA tag and nuclear localization signal | Nickase; generates 5' overhang; can induce NHEJ with paired offset gRNAs; reduced off-target effect. | NGG (preferred); NAG (less preferred) | Cell. 154: 1380 (2013) |
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| dCas9 | Catalytically inactive Cas9, a variant of Cas9 bearing both D10A and H840A mutations | Catalytically inactive but can be recruited by gRNAs to target sites; can be fused to other proteins to direct them to specific genomic locations (e.g. dCas9 fused with transcriptional activators or repressors can lead to activation or repression of target genes). | NGG (preferred); NAG (less preferred) | Nat Biotechnol. 32: 347 (2014) |
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| dCas9/VP64 | dCas9 fused with a VP64 acidic transactivation domain | Used for RNA-guided transcriptional activation | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 97:1495 (2000); Nat Methods. 10:973 (2013) |
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| dCas9/VPR | dCas9 fused with a tripartite VP64-p65-Rta (VPR) transactivation domain | Used for RNA-guided transcriptional activation. | NGG (preferred); NAG (less preferred) | Nat Methods. 12:326 (2015) |
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| dCas9/KRAB | dCas9 fused with a KRAB transcriptional silencing domain | Used for RNA-guided transcriptional repression | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 97:1495 (2000); Cell. 159:647 (2014) |
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| dCas9/KRAB/MeCP2 | dCas9 fused with a carboxy terminal bipartite repressor domain, KRAB-MeCP2 | Efficiently suppresses gene transcription by physically blocking RNA polymerase passage by dCas9 and further transcriptional inhibition by the KRAB-MeCP2 repressor domain. | NGG (preferred); NAG (less preferred) | Cell. 88:471 (1997); Nat Methods. 15:611 (2018) |
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| dCas9-10xGCN4_v4 | dCas9 with SV40 large T-antigen nuclear localization signals (nls) at both its amino and carboxyl termini, fused to an optimized GCN4 antibody-peptide pair (containing v4 version of the GCN4 peptide array with 10 copies of the peptide binding site, also know as SunTag10x_v4) | Modulates transcription of endogenous genes by recruitment of multiple copies of regulatory effector domains to genomic sites of interest targeted by dCas9 protein via binding of scFv antibody to a GCN4 repeating peptide array termed SunTag. | NGG (preferred); NAG (less preferred) | Cell. 159: 635 (2014) |
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| SpCas9-HF1 | High-fidelity variant of SpCas9 | Designed to reduce non-specific DNA contacts; fewer off-target cuts. | NGG (preferred); NAG (less preferred) | Nature. 529:490 (2016) |
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| eSpCas9 | Enhanced specificity SpCas9 variant | Low off-target effects and robust on-target cleavage. | NGG (preferred); NAG (less preferred) | Science. 351:84 (2016) |
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| BE3 | rAPOBEC–XTEN–Cas9_D10A–UGI (rat cytidine deaminase enzyme fused with Cas9_D10A and uracil DNA glycosylase inhibitor via an XTEN linker) | Converts cytosine (C) to uracil (U) between the 4th and 8th bases of the sgRNA binding site, targeting the nonbinding strand. The modified base can be permanently converted to thymine (T) following DNA replication or repair. | NGG (preferred); NAG (less preferred) | Nature. 533:420 (2016); Nat Methods. 14:710 (2017) |
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| Cas12a | Also known as Cpf1; single RNA-guided endonuclease of a class 2 CRISPR-Cas system from Acidaminococcus, human codon-optimized, and linked to an HA tag and nuclear localization signal | Cleaves DNA via a staggered DNA double strand break. Able to process its own CRISPR RNA (crRNA), which can simplify multiplexed genome editing. | TTTN | Cell. 163:759 (2015);Nat Biotechnol. 35:31 (2017) |
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| Cas9/mSA | hCas9 fused with monomeric streptavidin | Used for efficient generation of targeted large insertions using biotinylated repair templates. | NGG (preferred); NAG (less preferred) | Biotechnol Bioeng. 110:57 (2013); Nat Biotechnol. 36:632 (2018) |
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| SaCas9 | Staphylococcus aureus CRISPR associated protein 9 with HA tag and nuclear localization signal | Shorter sequence; especially useful in vector systems that have limited cargo space (e.g. AAV vectors). | NNGRR; NNGRRT(preferred) | Nature. 520:186 (2015) |
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| hCas9:T2A:Neo | hCas9 and neomycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to neomycin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| hCas9:T2A:Puro | hCas9 and puromycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to puromycin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| hCas9:T2A:Hygro | hCas9 and hygromycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to hygromycin B. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| hCas9:T2A:Bsd | hCas9 and blasticidin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to blasticidin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| hCas9:P2A:EGFP | hCas9 and EGFP linked by P2A | Allows cells to express hCas9 protein and be visualized by green fluorescence. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| hCas9:P2A:mCherry | hCas9 and mCherry linked by P2A | Allows cells to express hCas9 protein and be visualized by red fluorescence. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder |
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| DmCas9 | Fruit fly (Drosophila melanogaster) codon-optimized CRISPR associated protein 9 with nuclear localization signal | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 111:E2967 (2014) |
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| CjCas9 | A small Cas9 orthologue derived from Campylobacter jejuni with a carboxy terminal nuclear localization signal and HA tag | Shorter sequence; especially useful in vector systems that have limited cargo space (e.g. AAV vectors). | NNNNR(A/G)Y(C/T)AC; NNNNACAC (most preferred) | Nat Commun. 8:14500 (2017) |
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| CeCas9 | S. pyogenes Cas9 codon-optimized for C. elegans, containing multiple synthetic introns, a carboxy terminal nuclear localization signal, and an HA tag | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Nat Methods. 10:1028 (2013) |
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