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.
Thrombin R CRISPR/Cas9 KO Plasmid (m): sc-420264
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Thrombin R CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 Thrombin R 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: Thrombin R Antibody (ATAP2): sc-13503
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Thrombin R CRISPR/Cas9 KO Plasmid (m) | sc-420264 | 20 µg | $397.00 |
Overview
F2r encodes thrombin receptor (protease-activated receptor-1, PAR1), a GPCR activated by thrombin-dependent proteolytic cleavage that exposes a tethered ligand, initiating intracellular signaling through Gαq, Gα12/13, and Gαi pathways. In mouse cells, Thrombin R regulates platelet and endothelial responses, vascular tone, barrier function, and inflammatory signaling via downstream MAPK/ERK, RhoA/ROCK, PLCβ–Ca²⁺ mobilization, and NF-κB-associated transcriptional programs. F2r activity integrates coagulation with cellular responses to tissue injury and hemostatic stress, influencing thromboinflammatory crosstalk and vascular remodeling. Dysregulated PAR1 signaling has been linked to models of thrombosis, atherosclerosis, sepsis-associated vascular dysfunction, and tumor–stroma interactions in the microenvironment, making it relevant for mechanistic studies of vascular and inflammatory disease biology.
Thrombin R CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the F2r gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the F2r 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 F2r 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 Thrombin R protein expression.
This CRISPR knockout system enables efficient generation of F2r-deficient cell models for investigation of Thrombin R signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting F2r exon(s) critical for Thrombin R 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 F2r genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Thrombin R CRISPR/Cas9 KO Plasmid (m) and Thrombin R CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the F2r locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Thrombin R HDR Plasmid (m) and Thrombin R HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by F2r homology arms to support homology-directed repair at defined F2r 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.