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/Cas9 KO Plasmid (h): sc-401807
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Epac CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the Epac genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- 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/Cas9 KO Plasmid (h) | sc-401807 | 20 µg | $397.00 |
Overview
RAPGEF3 encodes Epac (exchange protein directly activated by cAMP), a guanine nucleotide exchange factor that links cAMP signaling to activation of Rap1 and Rap2 small GTPases. Epac modulates intracellular signal transduction in processes including cell adhesion, endothelial barrier regulation, secretion, and calcium-dependent responses through cAMP-dependent pathways that intersect with PKA, MAPK, and integrin signaling networks. In human cells, RAPGEF3/Epac activity influences vascular and immune cell functions and has been studied in contexts such as metabolic regulation, inflammatory signaling, and tumor-associated cell migration phenotypes. Dissecting RAPGEF3-dependent cAMP–Rap signaling helps clarify how second-messenger dynamics shape cytoskeletal remodeling and compartmentalized signaling complexes.
Epac CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RAPGEF3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RAPGEF3 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the RAPGEF3 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish Epac protein expression.
This CRISPR knockout system enables efficient generation of RAPGEF3-deficient cell models for investigation of Epac signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RAPGEF3 exon(s) critical for Epac function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple RAPGEF3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Epac CRISPR/Cas9 KO Plasmid (h) and Epac CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RAPGEF3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Epac HDR Plasmid (h) and Epac HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RAPGEF3 homology arms to support homology-directed repair at defined RAPGEF3 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.