Are all gRNA expression systems compatible with CRISPRko/CRISPRi/CRISPRa?
Assuming the (d)Cas9 is expressed separately, would one gRNA expression vector work for CRISPRko, CRISPRi, and CRISPRa?
Eg, can I use pLentiGuide for CRISPRko, CRISPRi, and CRISPRa assuming the cells have the appropriate (d)Cas9?
Technically yes, practically probably not, as the gRNA vectors often carry other elements and have recruitment loops to increase the modulation of gene expression.
pLX_311-Cas9: SV40 promoter expresses blasticidin resistance; EF1a promoter expresses SpCas9 (Addgene 96924).
dCAS-VP64_Blast (pXPR_109): EF1a promoter expresses dSpCas9-VP64 and 2A site provides blasticidin resistance (Addgene 61425).
pXPR_118: EF1a promoter expresses dSpCas9 and 2A site provides hygromycin resistance (Addgene 113667).
pXPR_121: SV40 promoter expresses blasticidin resistance; EF1a promoter expresses KRAB-dSpCas9 (Addgene 96918).
lentiGuide-Puro (pXPR_003): EF1a promoter expresses puromycin resistance; U6 promoter expresses customizable sgRNA element (Addgene 52963).
lentiCRISPRv2 (pXPR_023): EF1a promoter expresses SpCas9 and 2A site provides puromycin resistance; U6 promoter expresses customizable sgRNA element (Addgene 52961).
pXPR_050: EF1a promoter expresses puromycin resistance; U6 promoter expresses customizable sgRNA element with tracr-v2 (Addgene 96925).
pXPR_502: PGK promoter expresses PP7 tethered to P65-HSF1 and 2A site provides puromycin resistance; U6 promoter expresses customizable sgRNA element with tracr-v14, containing 2 MS2 and 2 PP7 stem loops (Addgene 96923)
https://www.nature.com/articles/s41467-018-07901-8