CPS2 CRISPR/Cas9 KO Plasmid (h): sc-403519
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CPS2 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 CPS2 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: CPS2 Antibody (F-6): sc-376072
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CPS2 CRISPR/Cas9 KO Plasmid (h) | sc-403519 | 20 µg | $397.00 |
Overview
CAD encodes a multifunctional cytosolic enzyme complex that catalyzes the first committed steps of de novo pyrimidine biosynthesis, with CPS2 (carbamoyl-phosphate synthetase 2) initiating conversion of glutamine and bicarbonate toward UMP production. This activity supports nucleotide pool homeostasis required for DNA replication, RNA transcription, and membrane phospholipid synthesis, linking CAD function to cell-cycle progression and metabolic adaptation. CAD is regulated by nutrient and growth signaling pathways, including allosteric control by PRPP and feedback inhibition by UTP, and is modulated by phosphorylation downstream of MAPK/mTOR signaling. Dysregulated pyrimidine metabolism and altered CAD activity have been associated with proliferative phenotypes and metabolic reprogramming observed in multiple disease contexts, making it a useful node for studying nucleotide stress responses and genome maintenance.
CPS2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CAD gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CAD 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 CAD 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 CPS2 protein expression.
This CRISPR knockout system enables efficient generation of CAD-deficient cell models for investigation of CPS2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CAD exon(s) critical for CPS2 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 CAD genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CPS2 CRISPR/Cas9 KO Plasmid (h) and CPS2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CAD locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CPS2 HDR Plasmid (h) and CPS2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CAD homology arms to support homology-directed repair at defined CAD 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.