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Jaenisch Lab CRISPR Plasmids Available from Addgene


CRISPR-on-logo

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 [48222] and Lifetechnologies Gateway Destination Vectors ) to adapt to different systems by LR clonase reaction.

Jaenisch-lab-activator-schematic

Additional Resources:

Additionally information can be found at www.crispr-on.org/

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 Aug. PubMed . PMC .

Individual plasmids can be ordered via the links below:

Table 1. dCas9 Activators sgRNA Dual Expression

ID Plasmid
48236 pAC2-dual-dCas9VP48-sgExpression : Dual expression construct expressing both dCas9VP48 and sgRNA from separate promoters. Add to Cart
48238 pAC152-dual-dCas9VP64-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters. Add to Cart
48239 pAC153-dual-dCas9VP96-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters. Add to Cart
48240 pAC154-dual-dCas9VP160-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters. Add to Cart

Table 2. pmax dCas9 Activator Expression Plasmids

ID Plasmid
48223 pAC91-pmax-dCas9VP64 : dCas9VP64 on pmax expression vector. Add to Cart
48224 pAC92-pmax-dCas9VP96 : dCas9VP96 on pmax expression vector. Add to Cart
48225 pAC93-pmax-dCas9VP160 : dCas9VP160 on pmax expression vector. Add to Cart
48226 pAC94-pmax-dCas9VP160-2A-puro : dCas9VP160-2A-puro (puro-selectable) on pmax expression vecor. Add to Cart

Table 3. dCas9 Activators Gateway Donors

ID Plasmid
48214 pAC1-pCR8-dCas9VP48 : dCas9VP48 on Gateway donor vector pCR8/GW/TOPO. Add to Cart
48218 pAC84-pCR8-dCas9 : dCas9 on Gateway donor vector pCR8/GW/TOPO. Add to Cart
48219 pAC147-pCR8-dCas9VP64 : dCas9VP64 on Gateway donor vector pCR8/GW/TOPO. Add to Cart
48220 pAC148-pCR8-dCas9VP96 : dCas9VP96 on Gateway donor vector pCR8/GW/TOPO. Add to Cart
48221 pAC149-pCR8-dCas9VP160 : dCas9VP160 on Gateway donor vector pCR8/GW/TOPO. Add to Cart

Table 4. Gateway Destination Vector

ID Plasmid
48222 pAC90-pmax-DEST : Gateway Destination vector derived from Clontech's pmax (CAGGS) expression vector. Add to Cart