TIM-3 Double Nickase Plasmid (h): sc-416365-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TIM-3 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
- TIM-3 Double Nickase Plasmid (h) and TIM-3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HAVCR2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TIM-3 Antibody (182H2B): sc-517632
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TIM-3 Double Nickase Plasmid (h) | sc-416365-NIC | 20 µg | $410.00 | |||
TIM-3 Double Nickase Plasmid (h2) | sc-416365-NIC-2 | 20 µg | $410.00 |
HAVCR2 encodes the human T cell immunoglobulin and mucin domain–containing protein 3 (TIM-3), an immunoregulatory receptor expressed on activated T cells, regulatory T cells, NK cells, and myeloid populations. TIM-3 binds ligands such as galectin-9, phosphatidylserine, CEACAM1, and HMGB1 to modulate T cell receptor signaling, immune synapse function, cytokine production, and cell fate decisions linked to exhaustion and tolerance. Through crosstalk with checkpoint networks and inflammatory signaling programs, TIM-3 influences antigen presentation and effector responses in chronic immune stimulation contexts. Dysregulated HAVCR2/TIM-3 activity has been associated with altered immune homeostasis in cancer immunology, chronic viral infection, and autoimmune inflammation, making it a useful node for mechanistic studies of immune regulation.
TIM-3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HAVCR2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HAVCR2. 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 HAVCR2 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 HAVCR2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.