LIGHT CRISPR/Cas9 KO Plasmid (m): sc-424539
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LIGHT 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 LIGHT 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 | |
LIGHT CRISPR/Cas9 KO Plasmid (m) | sc-424539 | 20 µg | $397.00 |
Overview
Mouse Tnfsf14 encodes LIGHT (TNFSF14), a TNF superfamily cytokine that functions as a membrane-bound and soluble ligand for HVEM (TNFRSF14) and LTβR, coordinating bidirectional signaling between immune and stromal compartments. LIGHT engagement activates NF-κB and MAPK pathways to modulate T cell costimulation, dendritic cell function, and lymphoid tissue organization, with downstream effects on cytokine production and leukocyte trafficking. Dysregulated LIGHT signaling has been implicated in inflammatory and autoimmune phenotypes as well as tumor–immune interactions, making it a useful node for studying immune regulation and tissue inflammation in mouse models. In addition, Tnfsf14 contributes to barrier and vascular responses through LTβR-dependent programs that shape local microenvironments.
LIGHT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tnfsf14 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tnfsf14 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 Tnfsf14 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 LIGHT protein expression.
This CRISPR knockout system enables efficient generation of Tnfsf14-deficient cell models for investigation of LIGHT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tnfsf14 exon(s) critical for LIGHT 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 Tnfsf14 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by LIGHT CRISPR/Cas9 KO Plasmid (m) and LIGHT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tnfsf14 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by LIGHT HDR Plasmid (m) and LIGHT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tnfsf14 homology arms to support homology-directed repair at defined Tnfsf14 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.