Lunatic Fringe CRISPR/Cas9 KO Plasmid (m): sc-421414
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Lunatic 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 Lunatic 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
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Lunatic Fringe Antibody (TT-07): sc-100756
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Lunatic Fringe CRISPR/Cas9 KO Plasmid (m) | sc-421414 | 20 µg | $397.00 |
Overview
Lfng encodes Lunatic Fringe, a Golgi-localized β1,3-N-acetylglucosaminyltransferase that modifies O-fucosylated epidermal growth factor–like repeats on NOTCH receptors. By altering Notch receptor–ligand interactions, Lunatic Fringe tunes Notch signal strength and oscillatory dynamics that govern somite segmentation, boundary formation, and cell fate decisions during mouse development. Lfng activity is linked to the segmentation clock and broader Notch-dependent programs influencing progenitor maintenance and differentiation across multiple tissues. Dysregulated LFNG/Notch modulation is relevant to developmental patterning defects and provides a mechanistic entry point for studying Notch pathway contributions to disease-associated phenotypes in model systems.
Lunatic Fringe CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Lfng gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Lfng 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 Lfng 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 Lunatic Fringe protein expression.
This CRISPR knockout system enables efficient generation of Lfng-deficient cell models for investigation of Lunatic Fringe signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Lfng exon(s) critical for Lunatic 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 Lfng genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Lunatic Fringe CRISPR/Cas9 KO Plasmid (m) and Lunatic Fringe CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Lfng locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Lunatic Fringe HDR Plasmid (m) and Lunatic Fringe HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Lfng homology arms to support homology-directed repair at defined Lfng 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.