HM74 CRISPR/Cas9 KO Plasmid (m): sc-429832
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HM74 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 HM74 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: HM74 Antibody (D-8): sc-377292
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HM74 CRISPR/Cas9 KO Plasmid (m) | sc-429832 | 20 µg | $397.00 |
Overview
Hcar2 encodes HM74 (also known as GPR109A), a Gi/o-coupled G protein–coupled receptor that responds to niacin and endogenous metabolites such as β-hydroxybutyrate to regulate intracellular cAMP signaling. In mouse immune and barrier tissues, HM74 influences chemotaxis, inflammatory mediator production, and cellular metabolic adaptation through cAMP-dependent pathways and downstream transcriptional programs. Receptor activity has been linked to regulation of macrophage and neutrophil responses, epithelial homeostasis, and lipid-associated signaling networks. Altered Hcar2/HM74 signaling is therefore relevant to models of inflammation, metabolic stress, and tissue injury where GPCR-mediated immunometabolic crosstalk is a key variable.
HM74 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Hcar2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Hcar2 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 Hcar2 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 HM74 protein expression.
This CRISPR knockout system enables efficient generation of Hcar2-deficient cell models for investigation of HM74 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Hcar2 exon(s) critical for HM74 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 Hcar2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HM74 CRISPR/Cas9 KO Plasmid (m) and HM74 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Hcar2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HM74 HDR Plasmid (m) and HM74 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Hcar2 homology arms to support homology-directed repair at defined Hcar2 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.