Table 3.
Name | Gene/insert | Selectable marker | Purpose | Reference | |
---|---|---|---|---|---|
1.CRISPR/Cas9 plasmids that create single stranded break (Mammalian) | pX335-U6-chimeric_BB-CBh-hSpCas9n (D10A) | Humanized S. pyogenes Cas9(D10A) nickase | None | A human codon-optimized SpCas9 nickase and chimeric guide RNA expression plasmid | [28] |
2.CRISPR/Cas9 plasmids that create double stranded break (Mammalian) | pX330-U6-chimeric_BB-CBh-hSpCas9 | humanized S. pyogenes Cas9 | None | A human codon-optimized SpCas9 and chimeric guide RNA expression plasmid |
[28] |
pSpCas9 (BB)-2A–Puro (PX459) V2.0 | hSpCas9-2A-Puro V2.0 (Synthetic) | Puromycin | Cas9 from S. pyogenes with 2A–Puro, and cloning backbone for sgRNA (V2.0) | [31] | |
3.CRISPR/Cas9 plasmids which activate gene expression(Mammalian) | SP-dCas9-VPR | SP-dCas9-VPR (Homo sapiens) | Neomycin (select with G418) | SP-dCas9 with VP64-p65-Rta (VPR) fused to its C-terminus; mammalian vector | [33] |
dCAS9-VP64_GFP | dCAS9(D10A, H840A)-VP64_2A_GFP (Synthetic) | GFP | Expresses dCas9-VP64 activator with 2A GFP | [34] | |
lenti dCAS-VP64_Blast | dCAS9(D10A, N863A)-VP64_2A_Blast (Synthetic) | Blasticidin | 3rd generation lenti vector encoding dCas9-VP64 with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) | [34] | |
4.CRISPR/Cas9 plasmids which repress gene expression(Mammalian) | pSLQ1658-dCas9-EGFP | dCas9 fuse to EGFP(Homo sapiens) | Puromycin | Template for NLS-dCas9-NLS-EGFP fusion protein for CRISPR imaging (the recipient vector can beTetON 3G promoter system) | [35] |
pLV hUbC-dCas9-T2A-GFP | Humanized dead Cas9T2A GFP (Other) | Zeocin | Co-expresses human optimized S. pyogenes dCas9 and GFP | [36] | |
pJMP1 | dCas9 (Other) | None | Bacillus subtilis dCas9 expression vector; integrates into lacA/ganA | [37] | |
5.CRISPR/Cas9 plasmid for tagging protein | Protein pFETch_NCoA2 |
Human | Donor plasmid pFETCh_NCoA2 |
Donor vector for 3’ FLAG tag of human NCoA2 | [38] |
pFETch_RAD21 | Human | pFETCh_RAD21 | Homology arms and 3X Flag with P2A Neofor 3′ tagging of human RAD21 | [38] |