HM74 Double Nickase Plasmid (m): sc-429832-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HM74 Double Nickase Plasmid (m) 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
- HM74 Double Nickase Plasmid (m) and HM74 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Hcar2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HM74 Antibody (D-8): sc-377292
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HM74 Double Nickase Plasmid (m) | sc-429832-NIC | 20 µg | $410.00 |
Mouse Hcar2 encodes the G protein–coupled receptor HM74 (also known as GPR109A), a high-affinity receptor for niacin and related metabolites that couples primarily to Gi signaling. HM74 activation reduces intracellular cAMP and engages downstream pathways that influence inflammatory tone, chemotaxis, and lipid handling, including modulation of NF-κB–linked transcriptional programs. The receptor is expressed in immune and barrier-associated cell types and has been used to study metabolite sensing at the interface of diet, microbiota-derived signals, and innate immune regulation. Altered HM74/Hcar2 signaling has been implicated in models of dyslipidemia, atherosclerosis-related inflammation, colitis-like pathology, and tumor-associated immune microenvironments, making it relevant for mechanistic studies of immunometabolism.
HM74 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Hcar2 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Hcar2. 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 Hcar2 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 Hcar2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.