Trypsin-4 CRISPR/Cas9 KO Plasmid (m): sc-423521
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Trypsin-4 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 Trypsin-4 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 | |
Trypsin-4 CRISPR/Cas9 KO Plasmid (m) | sc-423521 | 20 µg | $397.00 |
Overview
Mouse Try4 encodes trypsin-4, a secreted serine protease that cleaves peptide substrates after basic residues and contributes to extracellular proteolysis. By regulating protein turnover in the pericellular space, Try4 can influence protease-activated receptor signaling, processing of bioactive peptides, and remodeling of the extracellular matrix. Protease activity of trypsin family members intersects with inflammatory cascades and epithelial barrier dynamics through coordinated action with other serine proteases and endogenous inhibitors. Altered protease–inhibitor balance is frequently examined in models of tissue injury, inflammation, and tumor microenvironment remodeling, making Try4 a useful target for mechanistic studies.
Trypsin-4 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Try4 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Try4 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 Try4 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 Trypsin-4 protein expression.
This CRISPR knockout system enables efficient generation of Try4-deficient cell models for investigation of Trypsin-4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Try4 exon(s) critical for Trypsin-4 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 Try4 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Trypsin-4 CRISPR/Cas9 KO Plasmid (m) and Trypsin-4 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Try4 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Trypsin-4 HDR Plasmid (m) and Trypsin-4 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Try4 homology arms to support homology-directed repair at defined Try4 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.