mAChR M2 CRISPR/Cas9 KO Plasmid (h): sc-402394
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mAChR M2 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 mAChR M2 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
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: mAChR M2 Antibody (M2-2-B3): sc-33712
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mAChR M2 CRISPR/Cas9 KO Plasmid (h) | sc-402394 | 20 µg | $397.00 |
Overview
CHRM2 encodes the M2 subtype of muscarinic acetylcholine receptor (mAChR M2), a Gi/o-coupled GPCR that dampens adenylyl cyclase activity to reduce intracellular cAMP and downstream PKA signaling. mAChR M2 also modulates ion channel conductance, influencing membrane excitability and neurotransmitter release in neuronal circuits, and regulates chronotropic and inotropic responses in cardiomyocytes. Through these mechanisms, CHRM2 participates in cholinergic neurotransmission, autonomic control, and broader GPCR-dependent signaling networks. Dysregulated muscarinic signaling has been linked in research settings to cardiac electrophysiology phenotypes and neuropsychiatric and cognitive processes, making CHRM2 a useful node for pathway interrogation.
mAChR M2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CHRM2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CHRM2 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 CHRM2 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 mAChR M2 protein expression.
This CRISPR knockout system enables efficient generation of CHRM2-deficient cell models for investigation of mAChR M2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CHRM2 exon(s) critical for mAChR M2 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 CHRM2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by mAChR M2 CRISPR/Cas9 KO Plasmid (h) and mAChR M2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CHRM2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by mAChR M2 HDR Plasmid (h) and mAChR M2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CHRM2 homology arms to support homology-directed repair at defined CHRM2 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.