CH25H CRISPR/Cas9 KO Plasmid (m): sc-419641
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CH25H 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 CH25H 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: CH25H Antibody (1G8): sc-293256
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CH25H CRISPR/Cas9 KO Plasmid (m) | sc-419641 | 20 µg | $397.00 |
Overview
Ch25h encodes cholesterol 25-hydroxylase (CH25H), an endoplasmic reticulum–associated enzyme that converts cholesterol to 25-hydroxycholesterol, an oxysterol that modulates membrane lipid composition and transcriptional control of cholesterol homeostasis. Through crosstalk with sterol-regulated programs such as SREBP signaling and oxysterol-responsive nuclear receptors, CH25H influences lipid metabolism, innate immune signaling, and inflammatory gene expression. In mouse models, Ch25h activity has been linked to macrophage activation states, antiviral restriction pathways, and regulation of cytokine networks. Dysregulated oxysterol production is relevant to studies of metabolic inflammation and immune-mediated tissue remodeling, where altered cholesterol handling can shape cellular stress responses and signaling outputs.
CH25H CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ch25h gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ch25h 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 Ch25h 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 CH25H protein expression.
This CRISPR knockout system enables efficient generation of Ch25h-deficient cell models for investigation of CH25H signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ch25h exon(s) critical for CH25H 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 Ch25h genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CH25H CRISPR/Cas9 KO Plasmid (m) and CH25H CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ch25h locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CH25H HDR Plasmid (m) and CH25H HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ch25h homology arms to support homology-directed repair at defined Ch25h 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.