mAChR M1 Double Nickase Plasmid (h): sc-401453-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mAChR M1 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 M1 Double Nickase Plasmid (h) and mAChR M1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CHRM1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: mAChR M1 Antibody (G-9): sc-365966
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mAChR M1 Double Nickase Plasmid (h) | sc-401453-NIC | 20 µg | $410.00 | |||
mAChR M1 Double Nickase Plasmid (h2) | sc-401453-NIC-2 | 20 µg | $410.00 |
CHRM1 encodes the human muscarinic acetylcholine receptor M1 (mAChR M1), a G protein–coupled receptor that primarily signals through Gq/11 to activate phospholipase Cβ, elevate intracellular Ca²⁺, and stimulate PKC-dependent transcriptional programs. mAChR M1 modulates neuronal excitability, synaptic plasticity, and cholinergic regulation of cortical and hippocampal circuits, with additional roles in peripheral smooth muscle and secretory tissues. Downstream engagement of MAPK/ERK and PI3K-associated signaling links CHRM1 activity to changes in gene expression, cellular metabolism, and receptor desensitization/internalization dynamics. Dysregulated cholinergic signaling involving CHRM1 has been studied in the context of cognitive and neuropsychiatric phenotypes, as well as broader GPCR pathway alterations observed in disease-relevant cellular models.
mAChR M1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CHRM1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CHRM1. 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 CHRM1 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 CHRM1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.