Radical Fringe CRISPR/Cas9 KO Plasmid (m): sc-422659
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Radical Fringe 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 Radical Fringe 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Radical Fringe CRISPR/Cas9 KO Plasmid (m) | sc-422659 | 20 µg | $397.00 |
Overview
Rfng encodes Radical Fringe, a Golgi-localized β1,3-N-acetylglucosaminyltransferase that modifies O-fucose residues on EGF-like repeats of Notch receptors, thereby tuning ligand-dependent Notch signaling strength. Through context-dependent enhancement of Delta-like versus Jagged/Serrate interactions, Radical Fringe helps regulate cell fate decisions, boundary formation, and tissue patterning during development and in adult stem/progenitor compartments. Altered Fringe-mediated glycosylation can shift Notch pathway output and downstream transcriptional programs controlling proliferation and differentiation. Dysregulated Notch signaling is implicated across diverse disease-relevant processes, making Rfng a useful node for dissecting Notch-driven phenotypes in mouse cellular and in vivo models.
Radical Fringe CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rfng gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rfng 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 Rfng 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 Radical Fringe protein expression.
This CRISPR knockout system enables efficient generation of Rfng-deficient cell models for investigation of Radical Fringe signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rfng exon(s) critical for Radical Fringe 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 Rfng genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Radical Fringe CRISPR/Cas9 KO Plasmid (m) and Radical Fringe CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rfng locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Radical Fringe HDR Plasmid (m) and Radical Fringe HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rfng homology arms to support homology-directed repair at defined Rfng 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.