mAChR M5 Double Nickase Plasmid (h): sc-402943-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mAChR M5 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
- mAChR M5 Double Nickase Plasmid (h) and mAChR M5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CHRM5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mAChR M5 Double Nickase Plasmid (h) | sc-402943-NIC | 20 µg | $410.00 | |||
mAChR M5 Double Nickase Plasmid (h2) | sc-402943-NIC-2 | 20 µg | $410.00 |
CHRM5 encodes the human muscarinic acetylcholine receptor M5 (mAChR M5), a G protein–coupled receptor that primarily engages Gq/11 signaling to stimulate phospholipase C, inositol trisphosphate/diacylglycerol production, intracellular Ca²⁺ mobilization, and downstream PKC and MAPK pathway activity. Through these cascades, M5 can modulate neuronal excitability, synaptic signaling, and neurovascular responses, while also influencing secretory and smooth muscle processes in a cell-type-dependent manner. CHRM5 expression and cholinergic signaling dynamics are frequently examined in the context of dopaminergic circuit regulation, neuroinflammation, and behavioral phenotypes relevant to substance use and other neuropsychiatric research areas.
mAChR M5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CHRM5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CHRM5. 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 CHRM5 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 CHRM5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.