ECP CRISPR/Cas9 KO Plasmid (h): sc-404327
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ECP CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 ECP 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: ECP Antibody (C3): sc-517595
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ECP CRISPR/Cas9 KO Plasmid (h) | sc-404327 | 20 µg | $397.00 |
Overview
RNASE3 encodes eosinophil cationic protein (ECP), a granule-associated ribonuclease released by activated eosinophils during immune responses. ECP exhibits antimicrobial and cytotoxic activities and contributes to eosinophil-mediated tissue remodeling through interactions with cell membranes, extracellular matrix components, and innate immune signaling. RNASE3 expression and degranulation are linked to type 2 inflammation and eosinophilic responses that shape airway, skin, and gastrointestinal mucosal biology. Dysregulated ECP levels are frequently used as a molecular readout of eosinophil activation in inflammatory disease research, including asthma and other allergic disorders.
ECP CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RNASE3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RNASE3 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 RNASE3 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 ECP protein expression.
This CRISPR knockout system enables efficient generation of RNASE3-deficient cell models for investigation of ECP signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RNASE3 exon(s) critical for ECP 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 RNASE3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ECP CRISPR/Cas9 KO Plasmid (h) and ECP CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RNASE3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ECP HDR Plasmid (h) and ECP HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RNASE3 homology arms to support homology-directed repair at defined RNASE3 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.