Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
mHMGCS CRISPR/Cas9 KO Plasmid (h): sc-403941
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mHMGCS 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 mHMGCS 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: mHMGCS Antibody (B-8): sc-393256
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mHMGCS CRISPR/Cas9 KO Plasmid (h) | sc-403941 | 20 µg | $397.00 |
Overview
HMGCS2 encodes mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase (mHMGCS), the rate-limiting enzyme of ketogenesis that catalyzes formation of HMG-CoA from acetoacetyl-CoA and acetyl-CoA. It links hepatic fatty acid β-oxidation to production of ketone bodies during fasting and metabolic stress, influencing mitochondrial energy homeostasis and redox balance. HMGCS2 activity is integrated with PPARα-driven transcriptional programs and nutrient-sensing pathways that coordinate lipid utilization, gluconeogenic flux, and oxidative metabolism. Dysregulation of HMGCS2 has been associated with inborn errors of ketone body synthesis and with altered metabolic phenotypes observed in liver disease and cancer metabolism studies.
mHMGCS CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the HMGCS2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the HMGCS2 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 HMGCS2 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 mHMGCS protein expression.
This CRISPR knockout system enables efficient generation of HMGCS2-deficient cell models for investigation of mHMGCS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting HMGCS2 exon(s) critical for mHMGCS 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 HMGCS2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by mHMGCS CRISPR/Cas9 KO Plasmid (h) and mHMGCS CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the HMGCS2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by mHMGCS HDR Plasmid (h) and mHMGCS HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by HMGCS2 homology arms to support homology-directed repair at defined HMGCS2 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.