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.
phostensin CRISPR/Cas9 KO Plasmid (h): sc-407125
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- phostensin 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 phostensin 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: phostensin Antibody (H-9): sc-376816
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
phostensin CRISPR/Cas9 KO Plasmid (h) | sc-407125 | 20 µg | $397.00 |
Overview
PPP1R18 encodes phostensin, an actin-associated regulatory subunit that helps localize protein phosphatase 1 (PP1) activity to the cortical cytoskeleton. By coupling PP1 to filamentous actin, phostensin contributes to control of actin filament dynamics, cell shape, and adhesion-dependent signaling, with downstream effects on cell motility and immune cell function. This PP1-scaffolding role links PPP1R18 to phosphorylation-dependent remodeling processes that coordinate cytoskeletal organization and membrane-proximal signaling complexes. Dysregulation of phosphatase targeting and actin remodeling is frequently relevant to cancer cell migration and invasion phenotypes as well as inflammatory and immune-associated processes.
phostensin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PPP1R18 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PPP1R18 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 PPP1R18 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 phostensin protein expression.
This CRISPR knockout system enables efficient generation of PPP1R18-deficient cell models for investigation of phostensin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PPP1R18 exon(s) critical for phostensin 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 PPP1R18 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by phostensin CRISPR/Cas9 KO Plasmid (h) and phostensin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PPP1R18 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by phostensin HDR Plasmid (h) and phostensin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PPP1R18 homology arms to support homology-directed repair at defined PPP1R18 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.