TDAG8 CRISPR/Cas9 KO Plasmid (h): sc-416613
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TDAG8 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 TDAG8 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 | |
TDAG8 CRISPR/Cas9 KO Plasmid (h) | sc-416613 | 20 µg | $397.00 |
Overview
GPR65 encodes TDAG8, a proton-sensing G protein–coupled receptor that detects extracellular acidification and transduces signals through cAMP/PKA and downstream transcriptional programs. TDAG8 is enriched in immune and hematopoietic contexts, where it modulates inflammatory responses, chemotaxis, and cell survival in acidic microenvironments. By coupling extracellular pH to GPCR signaling, TDAG8 influences stress-response pathways and cytokine networks relevant to tissue inflammation and tumor-associated acidosis. Altered GPR65 activity has been studied in relation to immune dysregulation and microenvironment-driven phenotypes, supporting its use as a mechanistic node in GPCR and pH-sensing biology.
TDAG8 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GPR65 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GPR65 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 GPR65 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 TDAG8 protein expression.
This CRISPR knockout system enables efficient generation of GPR65-deficient cell models for investigation of TDAG8 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GPR65 exon(s) critical for TDAG8 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 GPR65 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TDAG8 CRISPR/Cas9 KO Plasmid (h) and TDAG8 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GPR65 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TDAG8 HDR Plasmid (h) and TDAG8 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GPR65 homology arms to support homology-directed repair at defined GPR65 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.