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.
A-Raf CRISPR/Cas9 KO Plasmid (h): sc-401799
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- A-Raf 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 A-Raf 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: A-Raf Antibody (A-5): sc-166771
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
A-Raf CRISPR/Cas9 KO Plasmid (h) | sc-401799 | 20 µg | $397.00 |
Overview
ARAF encodes A-Raf, a serine/threonine kinase of the RAF family that couples activated RAS at the plasma membrane to the MAPK/ERK signaling cascade. A-Raf participates in regulating mitogenic signaling, differentiation, and stress responses through RAF–MEK–ERK pathway control and crosstalk with other survival and apoptotic programs. Although BRAF and RAF1 are more frequently implicated, dysregulated RAF signaling and altered ARAF expression or function can contribute to aberrant pathway output relevant to oncogenic transformation and altered cell fate decisions. As a component of a central signal transduction module, A-Raf is studied in contexts such as RAS-driven signaling dynamics, feedback regulation, and pathway rewiring in disease-relevant cell models.
A-Raf CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ARAF gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ARAF 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 ARAF 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 A-Raf protein expression.
This CRISPR knockout system enables efficient generation of ARAF-deficient cell models for investigation of A-Raf signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ARAF exon(s) critical for A-Raf 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 ARAF genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by A-Raf CRISPR/Cas9 KO Plasmid (h) and A-Raf CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ARAF locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by A-Raf HDR Plasmid (h) and A-Raf HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ARAF homology arms to support homology-directed repair at defined ARAF 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.