Mast Cell Protease 5 CRISPR/Cas9 KO Plasmid (m): sc-421601
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mast Cell Protease 5 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 Mast Cell Protease 5 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 | |
Mast Cell Protease 5 CRISPR/Cas9 KO Plasmid (m) | sc-421601 | 20 µg | $397.00 |
Overview
Cma1 encodes mouse mast cell protease 5, a chymase-family serine protease stored in secretory granules of connective tissue–type mast cells and released upon FcεRI-driven degranulation. Mast cell protease 5 participates in extracellular matrix remodeling and regulation of vasoactive and proinflammatory mediators through proteolytic processing of peptides and cytokine-related substrates, shaping leukocyte recruitment and tissue responses. Through these activities it contributes to pathways underlying allergic inflammation, airway hyperreactivity, and tissue fibrosis, and it is frequently studied in models of cutaneous inflammation and cardiopulmonary remodeling. Altered mast cell protease 5 activity has been linked to changes in innate–adaptive immune crosstalk and barrier tissue homeostasis in inflammatory disease settings.
Mast Cell Protease 5 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cma1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cma1 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 Cma1 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 Mast Cell Protease 5 protein expression.
This CRISPR knockout system enables efficient generation of Cma1-deficient cell models for investigation of Mast Cell Protease 5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Cma1 exon(s) critical for Mast Cell Protease 5 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 Cma1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Mast Cell Protease 5 CRISPR/Cas9 KO Plasmid (m) and Mast Cell Protease 5 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cma1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Mast Cell Protease 5 HDR Plasmid (m) and Mast Cell Protease 5 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cma1 homology arms to support homology-directed repair at defined Cma1 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.