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.
PNP CRISPR/Cas9 KO Plasmid (h): sc-404914
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PNP 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 PNP 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: PNP Antibody (F-10): sc-365551
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PNP CRISPR/Cas9 KO Plasmid (h) | sc-404914 | 20 µg | $397.00 |
Overview
Purine nucleoside phosphorylase (PNP) is a cytosolic enzyme in the purine salvage pathway that catalyzes phosphorolysis of inosine and guanosine nucleosides to generate hypoxanthine or guanine, thereby supporting nucleotide homeostasis and recycling. By controlling the balance of purine metabolites, PNP influences cellular energy status, redox conditions, and DNA/RNA precursor availability, with downstream impacts on proliferation and stress responses. PNP activity is particularly important in immune cell metabolism, where purine flux modulates lymphocyte survival and function. Dysregulation or inherited deficiency of PNP is associated with immunodeficiency phenotypes and altered purine metabolite accumulation, making it a relevant target for mechanistic studies of metabolism-linked cell fate decisions.
PNP CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PNP gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PNP 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 PNP 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 PNP protein expression.
This CRISPR knockout system enables efficient generation of PNP-deficient cell models for investigation of PNP signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PNP exon(s) critical for PNP 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 PNP genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PNP CRISPR/Cas9 KO Plasmid (h) and PNP CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PNP locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PNP HDR Plasmid (h) and PNP HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PNP homology arms to support homology-directed repair at defined PNP 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.