CD154 Double Nickase Plasmid (h): sc-401682-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD154 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- CD154 Double Nickase Plasmid (h) and CD154 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CD40LG. One or both designs may be available
- 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 Double Nickase Plasmid (h) | sc-401682-NIC | 20 µg | $410.00 | |||
CD154 Double Nickase Plasmid (h2) | sc-401682-NIC-2 | 20 µg | $410.00 |
CD40LG encodes CD154 (CD40 ligand), a type II membrane protein transiently expressed on activated CD4+ T cells that engages CD40 on B cells, dendritic cells, and macrophages to coordinate adaptive and innate immune crosstalk. CD154–CD40 signaling promotes B-cell activation, germinal center formation, immunoglobulin class-switch recombination, and dendritic cell licensing through NF-κB and MAPK-driven transcriptional programs. This axis also shapes cytokine production and costimulatory signaling within T follicular helper responses and antigen presentation pathways. Dysregulated CD40LG activity or expression is associated with immune deficiency and immune hyperactivation phenotypes, making it relevant for mechanistic studies of humoral immunity, inflammation, and lymphoid microenvironment regulation.
CD154 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CD40LG locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CD40LG. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt CD40LG function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of CD40LG-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.