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.
RIP/Rab CRISPR/Cas9 KO Plasmid (m): sc-420946
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RIP/Rab 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 RIP/Rab 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: RIP/Rab Antibody (H-2): sc-166651
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RIP/Rab CRISPR/Cas9 KO Plasmid (m) | sc-420946 | 20 µg | $397.00 |
Overview
Agfg1 encodes RIP/Rab, an ArfGAP that couples ADP-ribosylation factor signaling to endocytic membrane traffic and cytoskeletal organization. Through regulation of small GTPase cycles, RIP/Rab contributes to receptor internalization, vesicle budding, and endosome-to-plasma-membrane recycling, processes that influence growth factor signaling outputs. Disruption of these trafficking nodes can alter cell polarity, migration, and stress responses, linking Agfg1-dependent pathways to phenotypes relevant to neurobiology and cancer cell behavior in model systems. In mouse studies, Agfg1 is therefore useful for probing how membrane trafficking intersects with signal transduction and cellular homeostasis.
RIP/Rab CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Agfg1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Agfg1 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 Agfg1 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 RIP/Rab protein expression.
This CRISPR knockout system enables efficient generation of Agfg1-deficient cell models for investigation of RIP/Rab signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Agfg1 exon(s) critical for RIP/Rab 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 Agfg1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RIP/Rab CRISPR/Cas9 KO Plasmid (m) and RIP/Rab CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Agfg1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RIP/Rab HDR Plasmid (m) and RIP/Rab HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Agfg1 homology arms to support homology-directed repair at defined Agfg1 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.