Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Epac CRISPR Activation Plasmid (h): sc-401807-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Epac CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Epac CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Epac CRISPR Activation Plasmid (h) and Epac CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RAPGEF3 transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Epac Antibody (A-5): sc-28366
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Epac CRISPR Activation Plasmid (h) | sc-401807-ACT | 20 µg | $397.00 | |||
Epac CRISPR Activation Plasmid (h2) | sc-401807-ACT-2 | 20 µg | $397.00 |
RAPGEF3 encodes Epac (exchange protein directly activated by cAMP), a guanine nucleotide exchange factor that links cAMP signaling to activation of the small GTPases Rap1 and Rap2. Through Rap-dependent control of integrin affinity, cell–cell junction stability, and cytoskeletal dynamics, Epac regulates adhesion, migration, barrier function, and vesicle trafficking across multiple cell types. Epac signaling intersects with PKA-independent cAMP pathways and integrates with MAPK and PI3K regulatory networks to shape proliferation and stress responses. Dysregulated RAPGEF3/Epac activity has been implicated in processes relevant to cancer cell invasion, cardiovascular and inflammatory phenotypes, and metabolic control, making it a useful node for mechanistic pathway studies.
Epac CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAPGEF3 expression without altering the underlying DNA sequence.
Epac CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAPGEF3 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the RAPGEF3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Epac expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAPGEF3 locus and enabling the study of Epac-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Epac pathway restoration in tumor cells with silenced or reduced RAPGEF3 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.