Ras-GRF1 CRISPR/Cas9 KO Plasmid (m): sc-422605
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ras-GRF1 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 Ras-GRF1 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: Ras-GRF1 Antibody (D-12): sc-377234
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ras-GRF1 CRISPR/Cas9 KO Plasmid (m) | sc-422605 | 20 µg | $397.00 |
Overview
Rasgrf1 encodes Ras-GRF1, a calcium/calmodulin-regulated guanine nucleotide exchange factor that activates Ras family GTPases and couples neuronal activity to downstream MAPK/ERK signaling. In mouse, Ras-GRF1 contributes to synaptic plasticity, learning and memory, and activity-dependent gene expression through modulation of Ras- and Rap-linked pathways, with additional roles in insulin/IGF-associated signaling and growth control in select tissues. Through these signaling nodes, Ras-GRF1 influences cellular proliferation, differentiation, and cytoskeletal dynamics, making it relevant to studies of neurodevelopmental phenotypes and dysregulated Ras pathway biology. Altered Ras-GRF1 function has been investigated in contexts of aberrant neuronal signaling and oncogenic pathway remodeling, supporting its use as a mechanistic entry point for pathway dissection.
Ras-GRF1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rasgrf1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rasgrf1 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 Rasgrf1 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 Ras-GRF1 protein expression.
This CRISPR knockout system enables efficient generation of Rasgrf1-deficient cell models for investigation of Ras-GRF1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rasgrf1 exon(s) critical for Ras-GRF1 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 Rasgrf1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ras-GRF1 CRISPR/Cas9 KO Plasmid (m) and Ras-GRF1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rasgrf1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ras-GRF1 HDR Plasmid (m) and Ras-GRF1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rasgrf1 homology arms to support homology-directed repair at defined Rasgrf1 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.