CD154 CRISPR/Cas9 KO Plasmid (m): sc-423452
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD154 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 CD154 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: CD154 Antibody (F-1): sc-374635
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD154 CRISPR/Cas9 KO Plasmid (m) | sc-423452 | 20 µg | $397.00 |
Overview
Cd40lg encodes CD154 (CD40 ligand), a type II membrane protein primarily expressed on activated T cells that engages CD40 on B cells, dendritic cells, and macrophages to coordinate adaptive immune responses. CD154–CD40 signaling drives NF-κB and MAPK pathway activation, promotes cytokine production, supports B cell proliferation and germinal center reactions, and contributes to immunoglobulin class switching. In mouse models, altered Cd40lg activity influences immune tolerance, antigen presentation, and inflammatory circuits relevant to autoimmunity, infection biology, and tumor immunology. Dissecting CD154-dependent costimulation is therefore central to studying T cell–APC crosstalk and lymphoid tissue organization.
CD154 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cd40lg gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cd40lg 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 Cd40lg 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 CD154 protein expression.
This CRISPR knockout system enables efficient generation of Cd40lg-deficient cell models for investigation of CD154 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Cd40lg exon(s) critical for CD154 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 Cd40lg genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CD154 CRISPR/Cas9 KO Plasmid (m) and CD154 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cd40lg locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CD154 HDR Plasmid (m) and CD154 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cd40lg homology arms to support homology-directed repair at defined Cd40lg 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.