cryptdin 25 CRISPR/Cas9 KO Plasmid (m): sc-419983
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- cryptdin 25 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 cryptdin 25 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 | |
cryptdin 25 CRISPR/Cas9 KO Plasmid (m) | sc-419983 | 20 µg | $397.00 |
Overview
Defa25 encodes cryptdin 25, a mouse Paneth cell–derived α-defensin that contributes to innate immunity at mucosal surfaces by exerting broad antimicrobial activity in the intestinal lumen. As a secreted cationic peptide, cryptdin 25 helps shape host–microbiota interactions and supports epithelial barrier homeostasis, linking antimicrobial effector functions to inflammatory signaling and microbial-sensing pathways. Altered defensin expression and Paneth cell dysfunction are frequently studied in the context of intestinal dysbiosis and chronic inflammatory states, where changes in antimicrobial peptide profiles can influence susceptibility to enteric infection and mucosal inflammation. Defa25 is therefore relevant for mechanistic studies of gut immunity, epithelial–microbe crosstalk, and lineage-specific secretory programs within the intestinal crypt.
cryptdin 25 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Defa25 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Defa25 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 Defa25 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 cryptdin 25 protein expression.
This CRISPR knockout system enables efficient generation of Defa25-deficient cell models for investigation of cryptdin 25 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Defa25 exon(s) critical for cryptdin 25 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 Defa25 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by cryptdin 25 CRISPR/Cas9 KO Plasmid (m) and cryptdin 25 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Defa25 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by cryptdin 25 HDR Plasmid (m) and cryptdin 25 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Defa25 homology arms to support homology-directed repair at defined Defa25 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.