ADF CRISPR/Cas9 KO Plasmid (h): sc-417540
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ADF 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 ADF 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 | |
ADF CRISPR/Cas9 KO Plasmid (h) | sc-417540 | 20 µg | $397.00 |
Overview
Human DSTN encodes actin depolymerizing factor (ADF), a core actin-binding protein that severs and depolymerizes F-actin to sustain filament turnover and maintain cytoskeletal homeostasis. By regulating actin dynamics, ADF supports processes including lamellipodia remodeling, endocytosis, cytokinesis, and cell migration, and it functionally intersects with Rho-family GTPase signaling and cofilin/ADF regulatory phosphorylation networks. Altered actin remodeling is linked to defects in epithelial barrier integrity, neuronal morphogenesis, and invasive cell behavior, making DSTN a relevant node for studying cytoskeleton-driven disease phenotypes. DSTN perturbation is therefore useful for dissecting how actin flux influences mechanotransduction, adhesion signaling, and stress responses in human cells.
ADF CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DSTN gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DSTN 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 DSTN 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 ADF protein expression.
This CRISPR knockout system enables efficient generation of DSTN-deficient cell models for investigation of ADF signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DSTN exon(s) critical for ADF 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 DSTN genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ADF CRISPR/Cas9 KO Plasmid (h) and ADF CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DSTN locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ADF HDR Plasmid (h) and ADF HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DSTN homology arms to support homology-directed repair at defined DSTN 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.