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.
PDI CRISPR/Cas9 KO Plasmid (m): sc-422099
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PDI 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 PDI 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: PDI Antibody (C-2): sc-74551
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PDI CRISPR/Cas9 KO Plasmid (m) | sc-422099 | 20 µg | $397.00 |
Overview
P4hb encodes protein disulfide isomerase (PDI), an abundant endoplasmic reticulum oxidoreductase that catalyzes disulfide bond formation, reduction, and isomerization to support folding and maturation of secretory and membrane proteins. PDI functions in ER proteostasis networks, partnering with chaperones and oxidoreductases to regulate oxidative protein folding and quality control, and it contributes to ER stress signaling and unfolded protein response pathways. Through its thiol-disulfide exchange activity and interactions with misfolded substrates, PDI influences redox homeostasis, protein trafficking, and cell survival decisions under proteotoxic stress. Dysregulated P4hb/PDI activity has been linked to pathological states characterized by chronic ER stress and altered redox balance, making it a relevant target for mechanistic studies in neurodegeneration, metabolic dysfunction, and cancer biology models.
PDI CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the P4hb gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the P4hb 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 P4hb 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 PDI protein expression.
This CRISPR knockout system enables efficient generation of P4hb-deficient cell models for investigation of PDI signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting P4hb exon(s) critical for PDI 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 P4hb genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PDI CRISPR/Cas9 KO Plasmid (m) and PDI CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the P4hb locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PDI HDR Plasmid (m) and PDI HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by P4hb homology arms to support homology-directed repair at defined P4hb 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.