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.
PIASx CRISPR/Cas9 KO Plasmid (m): sc-421655
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PIASx 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 PIASx 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: PIASx Antibody (D-12): sc-166494
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PIASx CRISPR/Cas9 KO Plasmid (m) | sc-421655 | 20 µg | $397.00 |
Overview
Pias2 encodes PIASx, a member of the protein inhibitor of activated STAT (PIAS) family that functions as a SUMO E3 ligase and transcriptional coregulator. PIASx modulates signal-dependent transcription by influencing STAT, SMAD, and nuclear receptor pathways, and it can alter chromatin-associated complexes through SUMOylation of target proteins. Through these activities, PIASx contributes to regulation of innate immune signaling, cytokine responses, and cell-cycle and differentiation programs. Dysregulation of PIASx-mediated SUMOylation and transcriptional control has been linked to aberrant inflammatory signaling and oncogenic transcriptional networks in mechanistic disease models.
PIASx CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pias2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pias2 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 Pias2 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 PIASx protein expression.
This CRISPR knockout system enables efficient generation of Pias2-deficient cell models for investigation of PIASx signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pias2 exon(s) critical for PIASx 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 Pias2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PIASx CRISPR/Cas9 KO Plasmid (m) and PIASx CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pias2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PIASx HDR Plasmid (m) and PIASx HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pias2 homology arms to support homology-directed repair at defined Pias2 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.