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.
ERp57 CRISPR/Cas9 KO Plasmid (m2): sc-420698-KO-2
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ERp57 CRISPR/Cas9 Knockout (KO) Plasmid (m2) 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 ERp57 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: ERp57 Antibody (MaP.ERp57): sc-23886
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ERp57 CRISPR/Cas9 KO Plasmid (m2) | sc-420698-KO-2 | 20 µg | $397.00 |
Overview
Pdia3 encodes ERp57, an endoplasmic reticulum protein disulfide isomerase that partners with calnexin and calreticulin to catalyze disulfide bond formation and isomerization during glycoprotein folding. ERp57 supports ER quality control and proteostasis by promoting maturation of secretory and membrane proteins and influencing unfolded protein response signaling under conditions of ER stress. In the immune system, ERp57 contributes to major histocompatibility complex class I peptide loading as part of the peptide-loading complex, linking redox-dependent folding to antigen presentation. Altered ERp57 activity has been associated with dysregulated protein homeostasis and oxidative stress pathways that are relevant to neurodegeneration, metabolic dysfunction, and cancer biology research.
ERp57 CRISPR/Cas9 KO Plasmid (m2) is a pool of plasmids designed for targeted disruption of the Pdia3 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pdia3 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 Pdia3 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 ERp57 protein expression.
This CRISPR knockout system enables efficient generation of Pdia3-deficient cell models for investigation of ERp57 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pdia3 exon(s) critical for ERp57 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 Pdia3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ERp57 CRISPR/Cas9 KO Plasmid (m) and ERp57 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pdia3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ERp57 HDR Plasmid (m) and ERp57 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pdia3 homology arms to support homology-directed repair at defined Pdia3 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.