Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
TLR4 Double Nickase Plasmid (h): sc-400068-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TLR4 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
- TLR4 Double Nickase Plasmid (h) and TLR4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TLR4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TLR4 Antibody (25): sc-293072
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TLR4 Double Nickase Plasmid (h) | sc-400068-NIC | 20 µg | $410.00 | |||
TLR4 Double Nickase Plasmid (h2) | sc-400068-NIC-2 | 20 µg | $410.00 |
Toll-like receptor 4 (TLR4) is a pattern-recognition receptor that detects bacterial lipopolysaccharide and select endogenous danger-associated ligands, initiating innate immune signaling in myeloid cells and barrier tissues. Ligand engagement promotes receptor complex formation with MD-2 and CD14 and activates MyD88- and TRIF-dependent cascades, driving NF-κB, AP-1, and IRF3/7 transcriptional programs and downstream cytokine and type I interferon responses. TLR4 signaling shapes macrophage polarization, inflammasome priming, and crosstalk with MAPK and JAK/STAT pathways that coordinate inflammatory homeostasis. Dysregulated TLR4 activity has been implicated in sepsis-associated inflammation, autoimmune and chronic inflammatory disorders, neuroinflammatory processes, and tumor microenvironment remodeling.
TLR4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TLR4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TLR4. 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 TLR4 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 TLR4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.