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.
PAR-4 CRISPR/Cas9 KO Plasmid (h): sc-401719
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PAR-4 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 PAR-4 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: PAR-4 Antibody (5F4): sc-293206
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PAR-4 CRISPR/Cas9 KO Plasmid (h) | sc-401719 | 20 µg | $397.00 |
Overview
F2RL3 encodes protease-activated receptor-4 (PAR-4), a thrombin-responsive G protein–coupled receptor that transduces extracellular protease signals into intracellular calcium mobilization and kinase cascades. PAR-4 couples to Gq and G12/13 pathways to regulate PLCβ signaling, RhoA-dependent cytoskeletal remodeling, and downstream MAPK and NF-κB programs that shape cellular activation states. In human platelets and vascular-associated cells, PAR-4 contributes to hemostatic and inflammatory signaling and interfaces with coagulation-linked pathways that influence cell adhesion and barrier responses. Altered F2RL3/PAR-4 activity has been investigated in contexts such as thrombosis-related biology, vascular inflammation, and tumor microenvironment–associated signaling where protease cues modulate proliferation and motility.
PAR-4 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the F2RL3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the F2RL3 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 F2RL3 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 PAR-4 protein expression.
This CRISPR knockout system enables efficient generation of F2RL3-deficient cell models for investigation of PAR-4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting F2RL3 exon(s) critical for PAR-4 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 F2RL3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PAR-4 CRISPR/Cas9 KO Plasmid (h) and PAR-4 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the F2RL3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PAR-4 HDR Plasmid (h) and PAR-4 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by F2RL3 homology arms to support homology-directed repair at defined F2RL3 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.