PP2Cκ CRISPR/Cas9 KO Plasmid (m): sc-433975
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PP2Cκ 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 PP2Cκ 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PP2Cκ CRISPR/Cas9 KO Plasmid (m) | sc-433975 | 20 µg | $397.00 |
Overview
Mouse Ppm1k encodes the mitochondrial protein phosphatase PP2Cκ, a Mg2+/Mn2+-dependent serine/threonine phosphatase that regulates branched-chain amino acid (BCAA) catabolism by dephosphorylating the branched-chain α-ketoacid dehydrogenase (BCKDH) complex. Through control of BCKDH activity, PP2Cκ influences mitochondrial nutrient flux, energy homeostasis, and metabolic signaling linked to amino acid availability. Disrupted Ppm1k function has been associated with altered BCAA levels and mitochondrial metabolic stress, making it relevant to studies of inborn errors of metabolism and broader metabolic phenotypes where BCAA handling is perturbed.
PP2Cκ CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ppm1k gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ppm1k 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 Ppm1k 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 PP2Cκ protein expression.
This CRISPR knockout system enables efficient generation of Ppm1k-deficient cell models for investigation of PP2Cκ signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ppm1k exon(s) critical for PP2Cκ 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 Ppm1k genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PP2Cκ CRISPR/Cas9 KO Plasmid (m) and PP2Cκ CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ppm1k locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PP2Cκ HDR Plasmid (m) and PP2Cκ HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ppm1k homology arms to support homology-directed repair at defined Ppm1k 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.