Ketohexokinase CRISPR/Cas9 KO Plasmid (m): sc-421261
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ketohexokinase 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 Ketohexokinase 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: Ketohexokinase Antibody (B-6): sc-377411
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ketohexokinase CRISPR/Cas9 KO Plasmid (m) | sc-421261 | 20 µg | $397.00 |
Overview
Khk encodes ketohexokinase (fructokinase), a key enzyme that catalyzes the phosphorylation of fructose to fructose-1-phosphate, committing fructose to downstream carbon flux. In mouse tissues with high fructose handling capacity, this activity interfaces with glycolysis, gluconeogenesis, and broader carbohydrate and energy metabolism through regulation of triose phosphate intermediates and hepatic metabolic signaling. Altered KHK-dependent fructose metabolism has been linked to metabolic stress phenotypes, including dysregulated lipid accumulation and insulin signaling, making Khk a useful node for studying nutrient sensing and metabolic remodeling. As part of fructose-responsive pathways, ketohexokinase perturbation can affect redox balance, ATP utilization, and transcriptional programs governing lipid and glucose homeostasis.
Ketohexokinase CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Khk gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Khk 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 Khk 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 Ketohexokinase protein expression.
This CRISPR knockout system enables efficient generation of Khk-deficient cell models for investigation of Ketohexokinase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Khk exon(s) critical for Ketohexokinase 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 Khk genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ketohexokinase CRISPR/Cas9 KO Plasmid (m) and Ketohexokinase CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Khk locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ketohexokinase HDR Plasmid (m) and Ketohexokinase HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Khk homology arms to support homology-directed repair at defined Khk 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.