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.
EAP30 CRISPR/Cas9 KO Plasmid (h): sc-406312
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EAP30 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 EAP30 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: EAP30 Antibody (C-11): sc-390747
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EAP30 CRISPR/Cas9 KO Plasmid (h) | sc-406312 | 20 µg | $397.00 |
Overview
SNF8 encodes EAP30, a core component of the ESCRT-II complex that coordinates endosomal sorting of ubiquitinated membrane proteins into multivesicular bodies for lysosomal degradation. Through ESCRT-dependent membrane remodeling, EAP30 supports receptor downregulation, endolysosomal trafficking, and coupling of ESCRT-II to ESCRT-III during intraluminal vesicle formation. These processes influence signaling pathway attenuation, membrane protein homeostasis, and aspects of cytokinesis and viral budding that rely on ESCRT machinery. Dysregulation of ESCRT-mediated trafficking has been linked to altered growth factor receptor signaling, proteostasis stress, and genomic instability, making SNF8/EAP30 relevant to studies of cancer biology and neurodegeneration-associated endolysosomal defects.
EAP30 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SNF8 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SNF8 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 SNF8 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 EAP30 protein expression.
This CRISPR knockout system enables efficient generation of SNF8-deficient cell models for investigation of EAP30 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SNF8 exon(s) critical for EAP30 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 SNF8 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by EAP30 CRISPR/Cas9 KO Plasmid (h) and EAP30 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SNF8 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by EAP30 HDR Plasmid (h) and EAP30 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SNF8 homology arms to support homology-directed repair at defined SNF8 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.