TDAG8 Double Nickase Plasmid (h): sc-416613-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TDAG8 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- TDAG8 Double Nickase Plasmid (h) and TDAG8 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GPR65. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TDAG8 Double Nickase Plasmid (h) | sc-416613-NIC | 20 µg | $410.00 | |||
TDAG8 Double Nickase Plasmid (h2) | sc-416613-NIC-2 | 20 µg | $410.00 |
Human GPR65 encodes TDAG8, a proton-sensing G protein–coupled receptor enriched in immune and hematopoietic contexts that couples primarily to Gs and cAMP signaling in response to extracellular acidosis. TDAG8 integrates pH-dependent cues within inflammatory microenvironments, influencing leukocyte activation, cytokine programs, and cell survival decisions, and intersects with broader GPCR-driven pathways that modulate MAPK and transcriptional responses. Altered regulation of GPR65 has been investigated in conditions characterized by tissue acidosis and dysregulated immune signaling, including inflammatory and autoimmune settings, as well as tumor microenvironment biology. These features make TDAG8 a useful entry point for studying how extracellular pH reshapes signaling networks and immune cell behavior.
TDAG8 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GPR65 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GPR65. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt GPR65 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of GPR65-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.