PYGL CRISPR/Cas9 KO Plasmid (m): sc-430989
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PYGL 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 PYGL 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: PYGL Antibody (C3): sc-517597
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PYGL CRISPR/Cas9 KO Plasmid (m) | sc-430989 | 20 µg | $397.00 |
Overview
Mouse Pygl encodes liver glycogen phosphorylase (PYGL), a rate-limiting enzyme that mobilizes glycogen by catalyzing phosphorolytic cleavage to generate glucose-1-phosphate for downstream glycolysis and glucose homeostasis. PYGL activity coordinates hepatic glycogenolysis with hormonal and nutrient-sensing inputs, linking carbohydrate availability to metabolic flux through central carbon pathways. Perturbation of PYGL function alters glycogen storage and utilization and is relevant to inborn errors of glycogen metabolism, making it a useful target for studying metabolic adaptation and energy balance in hepatocytes and whole-animal models. Pygl is therefore broadly informative for research on hepatic metabolic programming, fasting responses, and systems-level regulation of glucose metabolism.
PYGL CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pygl gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pygl 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 Pygl 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 PYGL protein expression.
This CRISPR knockout system enables efficient generation of Pygl-deficient cell models for investigation of PYGL signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pygl exon(s) critical for PYGL 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 Pygl genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PYGL CRISPR/Cas9 KO Plasmid (m) and PYGL CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pygl locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PYGL HDR Plasmid (m) and PYGL HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pygl homology arms to support homology-directed repair at defined Pygl 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.