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.
PCB CRISPR/Cas9 KO Plasmid (h): sc-401254
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PCB 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 PCB 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: PCB Antibody (D-9): sc-365673
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PCB CRISPR/Cas9 KO Plasmid (h) | sc-401254 | 20 µg | $397.00 |
Overview
Human PC (pyruvate carboxylase) encodes a biotin-dependent mitochondrial enzyme that catalyzes ATP-driven carboxylation of pyruvate to oxaloacetate, replenishing tricarboxylic acid (TCA) cycle intermediates and supporting anaplerosis. PC activity links central carbon metabolism to gluconeogenesis, lipogenesis, and amino acid biosynthesis, influencing mitochondrial redox balance and cellular energy homeostasis. Altered PC expression or function has been associated with inborn errors of metabolism and with metabolic reprogramming observed in several disease contexts, including cancer and liver disorders, where anaplerotic flux can affect proliferation and stress responses.
PCB CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PC gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PC 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 PC 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 PCB protein expression.
This CRISPR knockout system enables efficient generation of PC-deficient cell models for investigation of PCB signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PC exon(s) critical for PCB 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 PC genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PCB CRISPR/Cas9 KO Plasmid (h) and PCB CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PC locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PCB HDR Plasmid (h) and PCB HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PC homology arms to support homology-directed repair at defined PC 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.