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.
TLR5 Double Nickase Plasmid (h): sc-401154-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TLR5 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
- TLR5 Double Nickase Plasmid (h) and TLR5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TLR5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TLR5 Antibody (H-11): sc-518106
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TLR5 Double Nickase Plasmid (h) | sc-401154-NIC | 20 µg | $410.00 | |||
TLR5 Double Nickase Plasmid (h2) | sc-401154-NIC-2 | 20 µg | $410.00 |
Toll-like receptor 5 (TLR5) is a cell-surface pattern recognition receptor that detects bacterial flagellin and initiates innate immune signaling in epithelial cells and myeloid lineages. Upon ligand engagement, TLR5 signals primarily through MYD88 to activate IRAK–TRAF6 cascades, culminating in NF-κB and MAPK pathway activation and induction of pro-inflammatory cytokines and chemokines. TLR5 contributes to mucosal host defense and microbiome-immune homeostasis, and altered TLR5 activity has been linked to inflammatory disorders, infection susceptibility, and tumor-associated inflammation. Its well-defined ligand responsiveness makes TLR5 a useful node for dissecting PRR crosstalk, cytokine programs, and barrier immunity mechanisms.
TLR5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TLR5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TLR5. 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 TLR5 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 TLR5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.