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.
Rab 5A CRISPR/Cas9 KO Plasmid (m): sc-435445
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rab 5A 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 Rab 5A 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: Rab 5A Antibody (C-3): sc-515401
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rab 5A CRISPR/Cas9 KO Plasmid (m) | sc-435445 | 20 µg | $397.00 |
Overview
Rab5a encodes the small GTPase Rab 5A, a central regulator of early endosome dynamics that coordinates clathrin-mediated endocytosis, vesicle docking/fusion, and endosomal maturation through effectors such as EEA1 and PI3K signaling nodes. By controlling receptor internalization and trafficking, Rab 5A influences downstream signaling amplitude and duration for pathways including EGFR/MAPK and PI3K/AKT, and contributes to membrane recycling and cargo sorting toward lysosomal degradation. Rab 5A activity is also linked to cytoskeletal remodeling and neurite outgrowth via endosome–actin cross-talk. Dysregulated endocytic trafficking involving Rab5a has been implicated in neurodegeneration-related endosomal defects, altered immune receptor handling, and aberrant growth factor receptor signaling in proliferative disease models.
Rab 5A CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rab5a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rab5a 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 Rab5a 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 Rab 5A protein expression.
This CRISPR knockout system enables efficient generation of Rab5a-deficient cell models for investigation of Rab 5A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rab5a exon(s) critical for Rab 5A 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 Rab5a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Rab 5A CRISPR/Cas9 KO Plasmid (m) and Rab 5A CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rab5a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Rab 5A HDR Plasmid (m) and Rab 5A HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rab5a homology arms to support homology-directed repair at defined Rab5a 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.