stefin A2 CRISPR/Cas9 KO Plasmid (m): sc-423187
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- stefin A2 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 stefin A2 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
stefin A2 CRISPR/Cas9 KO Plasmid (m) | sc-423187 | 20 µg | $397.00 |
Overview
Stfa2 encodes the mouse cystatin-family protein stefin A2, a cytosolic inhibitor of papain-like cysteine proteases such as cathepsins. By restraining protease activity, stefin A2 contributes to regulation of intracellular proteolysis, lysosomal and endolysosomal protein turnover, and protection from protease-mediated damage during stress and inflammation. Altered balance between cysteine proteases and their inhibitors is relevant to pathways controlling antigen processing, epithelial barrier maintenance, and tissue remodeling. Dysregulation of this protease–inhibitor axis has been associated with inflammatory phenotypes and cancer-related processes in which cathepsin activity influences invasion and microenvironmental remodeling.
stefin A2 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Stfa2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Stfa2 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 Stfa2 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 stefin A2 protein expression.
This CRISPR knockout system enables efficient generation of Stfa2-deficient cell models for investigation of stefin A2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Stfa2 exon(s) critical for stefin A2 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 Stfa2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by stefin A2 CRISPR/Cas9 KO Plasmid (m) and stefin A2 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Stfa2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by stefin A2 HDR Plasmid (m) and stefin A2 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Stfa2 homology arms to support homology-directed repair at defined Stfa2 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.