TABLE 4.
The most commonly used plasmid vectors for expression of CRISPR/Cas9 targeting HBV genome.
| Name | Features | Promoter | Selection marker | Applications |
|---|---|---|---|---|
| pX330-U6-Chimeric BB-CBh-hSpCas9 | Single plasmid expressing both Cas9 and gRNA | U6 (gRNA), CBh (Cas9) | None intrinsic | Targeting HBV DNA for gene disruption and functional inactivation |
| pX458 (pSpCas9 BB-2A-GFP) | Cas9 and gRNA expression with GFP reporter | U6 (gRNA), CBh (Cas9) | GFP | Gene disruption of HBV DNA, monitoring transfection efficiency in HBV-infected cells |
| pX459 (pSpCas9 BB-2A-Puro) | Cas9 and gRNA expression with puromycin resistance | U6 (gRNA), CBh (Cas9) | Puromycin resistance | Gene disruption of HBV DNA, creation of stable cell lines resistant to HBV |
| pX601 (AAV-CMV: SaCas9-U6) | Smaller SaCas9 for AAV delivery | U6 (gRNA), CMV (SaCas9) | None intrinsic | In vivo editing of HBV DNA using AAV delivery systems |
| lentiCRISPR v2 | Lentiviral vector for stable integration and expression | U6 (gRNA), EF1α or CMV (Cas9) | Puromycin resistance | Long-term expression in HBV-infected cells, in vivo HBV models |
| pCW-Cas9 | Doxycycline-inducible Cas9 expression | U6 (gRNA), Tet (Cas9) | None intrinsic | Controlled activation of Cas9 nuclease in HBV gene editing, reducing off-target effects |
| pUC19-based gRNA and Cas9 Cloning Vectors | Simple vectors for cloning/expressing gRNAs and Cas9 | U6 (gRNA) | None intrinsic | Custom gRNA design targeting specific HBV sequences |
| pXPR_003 and pLX311-Cas9 (sgRNA/Cas9 Cloning Vectors) | High-throughput cloning vectors for gRNAs and Cas9 | U6 (gRNA) | None intrinsic | High-throughput screening of gRNAs targeting HBV |