mAChR M5 CRISPR/Cas9 KO Plasmid (h): sc-402943
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mAChR M5 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 M5 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mAChR M5 CRISPR/Cas9 KO Plasmid (h) | sc-402943 | 20 µg | $397.00 |
Overview
CHRM5 encodes the muscarinic acetylcholine receptor M5 (mAChR M5), a G protein–coupled receptor that primarily couples to Gq/11 to stimulate phospholipase C signaling, inositol phosphate production, intracellular Ca2+ mobilization, and PKC-dependent pathways. Through these second-messenger cascades, mAChR M5 modulates neuronal excitability, synaptic signaling, and vascular tone, and can engage MAPK/ERK signaling with downstream effects on gene expression programs. CHRM5 is expressed in discrete neural and peripheral cell populations, where cholinergic regulation intersects with dopaminergic circuitry and neurovascular regulation. Dysregulated muscarinic signaling has been implicated in neuropsychiatric and neurological phenotypes, supporting use of CHRM5 perturbation models to dissect cholinergic pathway contributions to disease-relevant cellular processes.
mAChR M5 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CHRM5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CHRM5 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 CHRM5 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 M5 protein expression.
This CRISPR knockout system enables efficient generation of CHRM5-deficient cell models for investigation of mAChR M5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CHRM5 exon(s) critical for mAChR M5 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 CHRM5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by mAChR M5 CRISPR/Cas9 KO Plasmid (h) and mAChR M5 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CHRM5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by mAChR M5 HDR Plasmid (h) and mAChR M5 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CHRM5 homology arms to support homology-directed repair at defined CHRM5 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.