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.
PCCB CRISPR/Cas9 KO Plasmid (m): sc-426258
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PCCB 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 PCCB 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: PCCB Antibody (F-6): sc-515740
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PCCB CRISPR/Cas9 KO Plasmid (m) | sc-426258 | 20 µg | $397.00 |
Overview
Mouse Pccb encodes the beta subunit of propionyl‑CoA carboxylase (PCCB), a biotin-dependent mitochondrial enzyme that catalyzes the conversion of propionyl‑CoA to methylmalonyl‑CoA. This reaction links branched-chain amino acid (valine, isoleucine), odd-chain fatty acid, and cholesterol side-chain catabolism to anaplerotic input into the TCA cycle via succinyl‑CoA. PCCB function supports mitochondrial metabolism and cellular energy homeostasis by preventing accumulation of propionyl‑CoA–derived metabolites. Disruption of propionyl‑CoA carboxylase activity is associated with propionic acidemia biology, making Pccb a useful locus for studying metabolic stress responses and mitochondrial pathway adaptation in mouse models.
PCCB CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pccb gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pccb 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 Pccb 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 PCCB protein expression.
This CRISPR knockout system enables efficient generation of Pccb-deficient cell models for investigation of PCCB signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pccb exon(s) critical for PCCB 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 Pccb genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PCCB CRISPR/Cas9 KO Plasmid (m) and PCCB CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pccb locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PCCB HDR Plasmid (m) and PCCB HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pccb homology arms to support homology-directed repair at defined Pccb 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.