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.
PKLR CRISPR/Cas9 KO Plasmid (m): sc-422273
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PKLR 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 PKLR 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: PKLR Antibody (E-2): sc-133222
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PKLR CRISPR/Cas9 KO Plasmid (m) | sc-422273 | 20 µg | $397.00 |
Overview
Mouse Pklr encodes pyruvate kinase liver and red blood cell (PKLR), a glycolytic enzyme that catalyzes conversion of phosphoenolpyruvate to pyruvate with concomitant ATP generation. PKLR activity helps control glycolytic flux, pyruvate availability for mitochondrial metabolism, and cellular energy balance, with particular importance in erythroid cells where glycolysis is a dominant ATP source. Regulation of PKLR intersects with nutrient-sensing and transcriptional programs that shape carbohydrate metabolism, including insulin-responsive and hypoxia-adaptive pathways. Altered PKLR function is associated with red blood cell metabolic dysfunction and related anemia phenotypes, making Pklr a useful node for studying metabolic stress responses and erythrocyte physiology in mouse models.
PKLR CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pklr gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pklr 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 Pklr 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 PKLR protein expression.
This CRISPR knockout system enables efficient generation of Pklr-deficient cell models for investigation of PKLR signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pklr exon(s) critical for PKLR 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 Pklr genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PKLR CRISPR/Cas9 KO Plasmid (m) and PKLR CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pklr locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PKLR HDR Plasmid (m) and PKLR HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pklr homology arms to support homology-directed repair at defined Pklr 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.