ITI-H5 Double Nickase Plasmid (h): sc-408967-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITI-H5 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
- ITI-H5 Double Nickase Plasmid (h) and ITI-H5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ITIH5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ITI-H5 Antibody (D-1): sc-390885
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ITI-H5 Double Nickase Plasmid (h) | sc-408967-NIC | 20 µg | $410.00 |
Inter-alpha-trypsin inhibitor heavy chain 5 (ITIH5) is a secreted extracellular matrix-associated protein that contributes to hyaluronan stabilization and matrix organization through covalent transfer of heavy chains to hyaluronan, influencing tissue hydration and biomechanical properties. In human cells, ITIH5 activity intersects with pathways that regulate cell–matrix adhesion, epithelial integrity, and stromal remodeling, affecting processes such as migration and invasion. Altered ITIH5 expression has been associated with tumor biology and metastatic potential in multiple tissue contexts, consistent with roles in extracellular matrix homeostasis and cell motility programs. These features make ITIH5 a useful target for dissecting how hyaluronan-rich matrices shape signaling cues and cellular behavior.
ITI-H5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ITIH5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ITIH5. 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 ITIH5 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 ITIH5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.