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.
PNPase CRISPR/Cas9 KO Plasmid (h): sc-404406
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PNPase 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 PNPase 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: PNPase Antibody (D-1): sc-271479
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PNPase CRISPR/Cas9 KO Plasmid (h) | sc-404406 | 20 µg | $397.00 |
Overview
PNPT1 encodes polyribonucleotide nucleotidyltransferase 1 (PNPase), a mitochondrial RNA-processing enzyme that participates in 3′–5′ exoribonuclease activity and RNA surveillance within the intermembrane space. PNPase contributes to mitochondrial gene expression by regulating the import and turnover of small RNAs and shaping mitoribosome function, thereby influencing oxidative phosphorylation capacity and cellular bioenergetics. Disruption of PNPT1 has been associated with mitochondrial dysfunction phenotypes, including defects in respiratory chain activity and altered stress responses linked to impaired mitochondrial RNA metabolism. As a result, PNPT1 is frequently studied in pathways connecting mitochondrial RNA homeostasis, innate immune signaling triggered by aberrant RNA species, and neurodevelopmental and metabolic disease-relevant mechanisms.
PNPase CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PNPT1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PNPT1 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 PNPT1 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 PNPase protein expression.
This CRISPR knockout system enables efficient generation of PNPT1-deficient cell models for investigation of PNPase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PNPT1 exon(s) critical for PNPase 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 PNPT1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PNPase CRISPR/Cas9 KO Plasmid (h) and PNPase CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PNPT1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PNPase HDR Plasmid (h) and PNPase HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PNPT1 homology arms to support homology-directed repair at defined PNPT1 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.