Ski CRISPR/Cas9 KO Plasmid (m): sc-422964
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ski 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 Ski 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: Ski Antibody (G8): sc-33693
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ski CRISPR/Cas9 KO Plasmid (m) | sc-422964 | 20 µg | $397.00 |
Overview
Ski encodes the Ski oncoprotein, a nuclear transcriptional coregulator that modulates gene expression programs controlling cell fate decisions. In mouse cells, Ski is best known for antagonizing TGF-β/SMAD signaling by interacting with SMAD complexes and corepressors, thereby shaping epithelial–mesenchymal transition, extracellular matrix deposition, and differentiation. Through these pathway effects, Ski influences proliferation, lineage commitment, and tissue remodeling, processes frequently perturbed in developmental abnormalities and cancer-associated phenotypes. Its regulatory roles make Ski a useful node for studying signal-dependent transcription, chromatin-associated repression, and context-specific growth control.
Ski CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ski gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ski 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 Ski 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 Ski protein expression.
This CRISPR knockout system enables efficient generation of Ski-deficient cell models for investigation of Ski signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ski exon(s) critical for Ski 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 Ski genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ski CRISPR/Cas9 KO Plasmid (m) and Ski CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ski locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ski HDR Plasmid (m) and Ski HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ski homology arms to support homology-directed repair at defined Ski 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.