Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Alix CRISPR/Cas9 KO Plasmid (m): sc-422154
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Alix 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 Alix 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: Alix Antibody (1A12): sc-53540
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Alix CRISPR/Cas9 KO Plasmid (m) | sc-422154 | 20 µg | $397.00 |
Overview
Pdcd6ip encodes ALIX, a multifunctional adaptor that couples membrane remodeling to endosomal trafficking through interactions with ESCRT components, including CHMP4 proteins, and partners such as TSG101. In mouse cells, ALIX contributes to multivesicular body biogenesis, exosome release, cytokinetic abscission, and plasma membrane repair, and it can influence apoptosis-linked signaling through its binding networks. ALIX-dependent ESCRT activity is also co-opted during budding of enveloped viruses and regulates turnover of membrane receptors, linking Pdcd6ip to pathways controlling cell signaling and proteostasis. Dysregulation of ESCRT/ALIX-mediated trafficking has been associated with neurodegeneration, tumor biology, and immune and viral pathogenesis in experimental systems, making Pdcd6ip a useful node for mechanistic studies of membrane dynamics.
Alix CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pdcd6ip gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pdcd6ip 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 Pdcd6ip 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 Alix protein expression.
This CRISPR knockout system enables efficient generation of Pdcd6ip-deficient cell models for investigation of Alix signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pdcd6ip exon(s) critical for Alix 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 Pdcd6ip genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Alix CRISPR/Cas9 KO Plasmid (m) and Alix CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pdcd6ip locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Alix HDR Plasmid (m) and Alix HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pdcd6ip homology arms to support homology-directed repair at defined Pdcd6ip 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.