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.
PFK-1 CRISPR/Cas9 KO Plasmid (h): sc-401023
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PFK-1 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 PFK-1 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: PFK-1 Antibody (E-4): sc-377346
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PFK-1 CRISPR/Cas9 KO Plasmid (h) | sc-401023 | 20 µg | $397.00 |
Overview
PFKM encodes the muscle isoform of phosphofructokinase-1 (PFK-1), a rate-limiting enzyme of glycolysis that catalyzes the ATP-dependent conversion of fructose-6-phosphate to fructose-1,6-bisphosphate. As a key control point for glycolytic flux, PFK-1 integrates allosteric regulation by cellular energy status and metabolites to coordinate glucose utilization with ATP demand. PFKM activity influences central carbon metabolism, lactate production, and downstream biosynthetic pathways coupled to proliferative and stress-adaptive programs. Genetic disruption of PFKM is linked to glycogen storage disease type VII (Tarui disease) and is used to probe how altered glycolysis contributes to metabolic dysfunction and cellular phenotypes relevant to neuromuscular and hematologic biology.
PFK-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PFKM gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PFKM 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 PFKM 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 PFK-1 protein expression.
This CRISPR knockout system enables efficient generation of PFKM-deficient cell models for investigation of PFK-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PFKM exon(s) critical for PFK-1 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 PFKM genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PFK-1 CRISPR/Cas9 KO Plasmid (h) and PFK-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PFKM locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PFK-1 HDR Plasmid (h) and PFK-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PFKM homology arms to support homology-directed repair at defined PFKM 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.