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.
Emt Double Nickase Plasmid (h): sc-402628-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Emt 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
- Emt Double Nickase Plasmid (h) and Emt Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ITK. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Emt Antibody (2F12): sc-23902
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Emt Double Nickase Plasmid (h) | sc-402628-NIC | 20 µg | $410.00 | |||
Emt Double Nickase Plasmid (h2) | sc-402628-NIC-2 | 20 µg | $410.00 |
Human ITK (IL2-inducible T-cell kinase) encodes a Tec family non-receptor tyrosine kinase that propagates signaling downstream of the T-cell receptor, co-stimulatory receptors, and chemokine inputs to shape T-cell activation, differentiation, and cytokine production. ITK integrates proximal phosphorylation events with PLCγ1 activation, calcium flux, MAPK signaling, and NFAT/NF-κB/AP-1–dependent transcriptional programs, influencing immune synapse formation and cytoskeletal remodeling. Dysregulated ITK signaling has been linked to altered Th2/Th17 polarization, impaired antiviral immunity, and aberrant lymphocyte activation states relevant to immunodeficiency and lymphoproliferative disease biology. As a pathway node connecting antigen recognition to transcriptional outputs, ITK is frequently studied in immune signaling networks, T-cell lineage decisions, and inflammatory signaling crosstalk.
Emt Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ITK locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ITK. 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 ITK 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 ITK-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.