TEM5 CRISPR/Cas9 KO Plasmid (h): sc-407178
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TEM5 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 TEM5 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TEM5 CRISPR/Cas9 KO Plasmid (h) | sc-407178 | 20 µg | $397.00 |
Overview
ADGRA2 (TEM5) encodes an adhesion G protein–coupled receptor with a large extracellular region that supports cell–cell and cell–matrix interactions and links extracellular cues to intracellular signaling. As a member of the adhesion GPCR family, TEM5 is associated with processes such as endothelial cell behavior, vascular and tissue remodeling, and regulation of cytoskeletal organization through GPCR-linked pathways. Expression of TEM5 has been reported in contexts relevant to tumor-associated vasculature and stromal remodeling, making it of interest for studies of angiogenic signaling and microenvironmental regulation. Dissecting ADGRA2 function can help clarify how adhesion GPCR signaling integrates extracellular adhesion dynamics with transcriptional and cytoskeletal responses in human cells.
TEM5 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ADGRA2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ADGRA2 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 ADGRA2 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 TEM5 protein expression.
This CRISPR knockout system enables efficient generation of ADGRA2-deficient cell models for investigation of TEM5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ADGRA2 exon(s) critical for TEM5 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 ADGRA2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TEM5 CRISPR/Cas9 KO Plasmid (h) and TEM5 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ADGRA2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TEM5 HDR Plasmid (h) and TEM5 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ADGRA2 homology arms to support homology-directed repair at defined ADGRA2 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.