Strumpellin CRISPR/Cas9 KO Plasmid (h): sc-402808
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Strumpellin 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 Strumpellin 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: Strumpellin Antibody (B-10): sc-377146
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Strumpellin CRISPR/Cas9 KO Plasmid (h) | sc-402808 | 20 µg | $397.00 |
Overview
WASHC5 encodes strumpellin, an essential core component of the WASH complex that regulates Arp2/3-dependent actin polymerization on endosomal membranes. Through coordination of endosomal tubulation and fission, strumpellin supports retromer-associated cargo sorting and recycling, influencing the trafficking of membrane receptors and nutrient transporters. WASHC5 activity integrates cytoskeletal remodeling with endosome-to-plasma membrane and endosome-to-Golgi pathways, shaping cell migration, signaling output, and lysosomal homeostasis. Pathogenic variation in WASHC5 has been linked to neurodegenerative phenotypes, including hereditary spastic paraplegia, highlighting its relevance to studies of neuronal maintenance and membrane trafficking stress.
Strumpellin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the WASHC5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the WASHC5 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 WASHC5 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 Strumpellin protein expression.
This CRISPR knockout system enables efficient generation of WASHC5-deficient cell models for investigation of Strumpellin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting WASHC5 exon(s) critical for Strumpellin 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 WASHC5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Strumpellin CRISPR/Cas9 KO Plasmid (h) and Strumpellin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the WASHC5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Strumpellin HDR Plasmid (h) and Strumpellin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by WASHC5 homology arms to support homology-directed repair at defined WASHC5 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.