PNUTS CRISPR/Cas9 KO Plasmid (m): sc-424580
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PNUTS 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 PNUTS 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: PNUTS Antibody (F-8): sc-271681
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PNUTS CRISPR/Cas9 KO Plasmid (m) | sc-424580 | 20 µg | $397.00 |
Overview
Mouse Ppp1r10 encodes PNUTS (protein phosphatase 1 regulatory subunit 10), a nuclear scaffold that targets PP1 to chromatin and RNA-associated complexes to coordinate phosphorylation-dependent control of gene expression. PNUTS participates in transcriptional regulation, DNA damage response signaling, and cell-cycle checkpoint control, influencing processes such as replication stress management and maintenance of genome stability. Through interactions with factors involved in chromatin organization and RNA processing, PNUTS helps couple phosphorylation dynamics to transcription elongation and RNA biogenesis. Dysregulation of PP1–PNUTS signaling has been linked to altered proliferative programs and genome instability phenotypes that are relevant to mechanistic studies of cancer-associated pathways and neurobiology.
PNUTS CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ppp1r10 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ppp1r10 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 Ppp1r10 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 PNUTS protein expression.
This CRISPR knockout system enables efficient generation of Ppp1r10-deficient cell models for investigation of PNUTS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ppp1r10 exon(s) critical for PNUTS 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 Ppp1r10 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PNUTS CRISPR/Cas9 KO Plasmid (m) and PNUTS CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ppp1r10 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PNUTS HDR Plasmid (m) and PNUTS HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ppp1r10 homology arms to support homology-directed repair at defined Ppp1r10 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.