PNGase CRISPR/Cas9 KO Plasmid (m): sc-425553
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PNGase 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 PNGase 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PNGase CRISPR/Cas9 KO Plasmid (m) | sc-425553 | 20 µg | $397.00 |
Overview
Mouse Ngly1 encodes peptide:N-glycanase (PNGase), a cytosolic deglycosylating enzyme that removes N-linked glycans from misfolded glycoproteins retrotranslocated from the endoplasmic reticulum. This activity is integral to ER-associated degradation (ERAD), coupling deglycosylation to ubiquitin–proteasome turnover and shaping proteostasis under unfolded protein response stress. NGLY1 also participates in regulation of NRF1/NFE2L1 processing, linking deglycosylation to cellular adaptation programs that control proteasome gene expression and stress resilience. Disruption of N-glycanase function is broadly relevant to mechanisms of proteotoxic stress, protein quality control, and neurodevelopmental phenotypes observed in NGLY1-related biology.
PNGase CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ngly1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ngly1 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 Ngly1 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 PNGase protein expression.
This CRISPR knockout system enables efficient generation of Ngly1-deficient cell models for investigation of PNGase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ngly1 exon(s) critical for PNGase 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 Ngly1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PNGase CRISPR/Cas9 KO Plasmid (m) and PNGase CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ngly1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PNGase HDR Plasmid (m) and PNGase HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ngly1 homology arms to support homology-directed repair at defined Ngly1 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.