TNFβ CRISPR/Cas9 KO Plasmid (m): sc-421477
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TNFβ 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 TNFβ 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: TNFβ Antibody (E-6): sc-28345
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TNFβ CRISPR/Cas9 KO Plasmid (m) | sc-421477 | 20 µg | $397.00 |
Overview
Lta encodes tumor necrosis factor beta (TNFβ, also known as lymphotoxin-α), a TNF superfamily cytokine primarily produced by activated lymphocytes that signals through TNFR1/TNFR2 and contributes to lymphoid organogenesis and inflammatory crosstalk. TNFβ activates canonical NF-κB and MAPK signaling to regulate leukocyte survival, cytokine networks, and adhesion molecule expression, shaping immune cell trafficking and tissue remodeling. In mouse systems, Lta function is tightly linked to organization of secondary lymphoid structures and coordination of adaptive immune responses. Dysregulated TNFβ signaling is frequently examined in models of chronic inflammation, autoimmunity, and tumor-associated immune microenvironments where stromal–immune interactions are altered.
TNFβ CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Lta gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Lta 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 Lta 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 TNFβ protein expression.
This CRISPR knockout system enables efficient generation of Lta-deficient cell models for investigation of TNFβ signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Lta exon(s) critical for TNFβ 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 Lta genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TNFβ CRISPR/Cas9 KO Plasmid (m) and TNFβ CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Lta locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TNFβ HDR Plasmid (m) and TNFβ HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Lta homology arms to support homology-directed repair at defined Lta 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.