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Table 3 Selective CRISPR/Cas9 plasmids

From: CRISPR/Cas9: the Jedi against the dark empire of diseases

  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]