Jaenisch Lab CRISPR Plasmids Available from Addgene
CRISPR-on is an RNA-guided transcriptional activator system with two components, the dCas9 activator protein and the single guide RNA (sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component provides the specificity and is designed to target promoters or enhancers of genes of interest.
Different flavors of activators were constructed, with VP48 containing 3x minimal VP16 domains, VP64 with 4, VP96 with 6, and VP160 with 10.
dCas9 activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression vector or the linear DNA containing the sgRNA expressing fragment produced by PCR), or expressed from a single dual-expression vector [48236~48240].
dCas9 activator ORFs (with Stop codon) are also available as Gateway donor vectors [48218~48221] and can be transferred to Gateway destination vectors (e.g., pmax-DEST  and (Link opens in a new window) Lifetechnologies Gateway Destination Vectors) to adapt to different systems by LR clonase reaction.
These plasmids are described in:
Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cheng AW, Wang H, Yang H, Shi L, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Cell Res. 2013 Oct;23(10):1163-71. doi: 10.1038/cr.2013.122. Epub 2013 Aug 27. (Link opens in a new window) PubMed (Link opens in a new window) Article.
Individual plasmids can be ordered via the links below:
Table 1. dCas9 Activators sgRNA Dual Expression
|48236||pAC2-dual-dCas9VP48-sgExpression: Dual expression construct expressing both dCas9VP48 and sgRNA from separate promoters.|
|48238||pAC152-dual-dCas9VP64-sgExpression: Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters.|
|48239||pAC153-dual-dCas9VP96-sgExpression: Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters.|
|48240||pAC154-dual-dCas9VP160-sgExpression: Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters.|
Table 2. pmax dCas9 Activator Expression Plasmids
|48223||pAC91-pmax-dCas9VP64: dCas9VP64 on pmax expression vector.|
|48224||pAC92-pmax-dCas9VP96: dCas9VP96 on pmax expression vector.|
|48225||pAC93-pmax-dCas9VP160: dCas9VP160 on pmax expression vector.|
|48226||pAC94-pmax-dCas9VP160-2A-puro: dCas9VP160-2A-puro (puro-selectable) on pmax expression vecor.|
Table 3. dCas9 Activators Gateway Donors
|48214||pAC1-pCR8-dCas9VP48: dCas9VP48 on Gateway donor vector pCR8/GW/TOPO.|
|48218||pAC84-pCR8-dCas9: dCas9 on Gateway donor vector pCR8/GW/TOPO.|
|48219||pAC147-pCR8-dCas9VP64: dCas9VP64 on Gateway donor vector pCR8/GW/TOPO.|
|48220||pAC148-pCR8-dCas9VP96: dCas9VP96 on Gateway donor vector pCR8/GW/TOPO.|
|48221||pAC149-pCR8-dCas9VP160: dCas9VP160 on Gateway donor vector pCR8/GW/TOPO.|
Table 4. Gateway Destination Vector
|48222||pAC90-pmax-DEST: Gateway Destination vector derived from Clontech's pmax (CAGGS) expression vector.|