CRISPR-Cas9 Plasmids: A Comprehensive Collection for Gene Editing Research

CRISPR-Cas9 is one of the most widely used genome-editing technologies for targeted gene knockout, knock-in, gene regulation, and functional genomics research. Different CRISPR plasmid systems have been developed for different organisms and applications, including conventional Cas9 systems, high-fidelity Cas9 variants, dCas9-based transcriptional regulation systems, and Cas12a (Cpf1)-based systems.

The following is a collection of commonly used CRISPR-related plasmids organized by research system.

Mammalian Cells

SpCas9 and Related CRISPR Plasmids

• pX260

• pX330

• pX334

• pX335

• pX458

• pX459

• pX459 V2.0

• pX461

• pX462

• pX462 V2

• pX551

• pX552

• pX601

• pX602

• pX330A-1x2

• pX330A-1x3

• pX330A_FokI-1x3

• pX330S-2-PITCh

• pX330A-FBL/PITCh

Lentiviral CRISPR Plasmids

• LentiCRISPR V1

• pLentiCRISPR V2

• LentiGuide-Puro

• LentiCas9-Blast

• pLentiCRISPR-E

• pLenti dCas VP64-Blast

Humanized Cas9 and sgRNA Systems

• pdCas9-humanized

• pgRNA-humanized

Additional CRISPR/Cas Systems

• pLSC-5

• pCW-Cas9

• pCDNA3.1-hLbCpf1

• pCDNA3.1-hLb2Cpf1

• pCDNA3.1-hAsCpf1

• pCDNA3.1-hFnCpf1

The Cpf1 systems listed above are based on Cas12a, an RNA-guided nuclease that differs from SpCas9 in PAM requirements, guide RNA architecture, and cleavage characteristics.

SaCas9 and Cas9 Variants

• BPK2101-saCas9-gRNA

• BPK2139-SaCas9

• MSP1830-SaCas9KKH

• MSP1101-SpCas9VRER

• MSP469-Cas9 VQR

• pSaCas9_GFP

• eSpCas9(1.1)

• peSpCas9-PURO

• peSpCas9-GFP

Additional Cas9 and sgRNA Vectors

• VVT1-gRNA

• pLX-sgRNA

• pCRIS-PITChv2-FBL

• hCas9

• hCas9_D10A

• pCS2-nCas9n

dCas9-Based Gene Regulation Plasmids

Catalytically inactive Cas9 (dCas9) can be fused with transcriptional activators, repressors, or epigenetic regulators for applications such as CRISPR activation (CRISPRa), CRISPR interference (CRISPRi), and targeted epigenome editing.

• pcDNA-dCas9

• pcDNA-dCas9-p300 Core

• pcDNA-dCas9-VP64

• pSQT1313

• pSQT1601

• pAC152-dual-dCas9VP64-sgExpression

• pHR-SFFV-dCas9-BFP-KRAB

• pHRdSV40-dCas9-10XGCN4_V4-P2A-BFP

• pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE

• pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS

• pHAGE-TO-dCas9

CRISPR Donor and Other Specialized Plasmids

• pCRISPRainbow-DONOR1

Bacterial Systems

CRISPR systems in bacteria can be used for genome editing, gene regulation, and functional genomic studies. Common bacterial CRISPR plasmids include:

• pCas

• pCas9

• pCRISPR

• pTargetF

• pET-NLS-Cas9-6×His

• pET22b-SaCas9

• pdCas9-Bacteria

• pwtCas9-Bacteria

• pgRNA-Bacteria

• pMJ806-MBP-cas9

• pY001 (pFnCpf1_full)

• pC001-huLshC2C2-MBP

• pC002-pACYC184-LshC2C2

• pC003-pACYC184-LshC2C2

• pET302-6HIS-dCas9-Halo

Plant Systems

CRISPR-based genome editing has been widely applied to plant functional genomics, gene knockout, gene insertion, and crop improvement research.

Commonly used plant CRISPR plasmids include:

• pHSN401

• pRGEB31

C. elegans

CRISPR-Cas9 has also been adapted for genome editing in Caenorhabditis elegans.

• pDD162 (Peft-3Cas9 + Empty sgRNA)

This vector provides Cas9 expression together with an sgRNA cloning framework for targeted genome editing in C. elegans.

Yeast

CRISPR-based systems have been developed for genome editing and genetic manipulation in yeast.

Commonly used plasmids include:

• pCRCT

• pCRISPRyl

• pCAS

Toxoplasma gondii

CRISPR-Cas9 systems can also be used for targeted genetic manipulation in Toxoplasma gondii.

• pSAG1::CAS9-U6::sgUPRT

This vector combines Cas9 expression driven by the SAG1 promoter with U6 promoter-driven sgRNA expression targeting the UPRT locus.

Summary

CRISPR plasmids can be broadly classified according to the Cas nuclease, delivery system, host organism, and intended application. Conventional Cas9 systems are commonly used for targeted gene knockout and genome editing, while dCas9-based systems enable transcriptional activation or repression without introducing double-strand DNA breaks.

For specific applications, factors such as host species, Cas protein selection, PAM requirements, sgRNA design, delivery method, selection marker, and whether permanent genome modification or transcriptional regulation is desired should be considered when selecting an appropriate CRISPR plasmid.