Pre-TCRα Double Nickase Plasmid (h): sc-417210-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pre-TCRα 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
- Pre-TCRα Double Nickase Plasmid (h) and Pre-TCRα Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PTCRA. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pre-TCRα Double Nickase Plasmid (h) | sc-417210-NIC | 20 µg | $410.00 | |||
Pre-TCRα Double Nickase Plasmid (h2) | sc-417210-NIC-2 | 20 µg | $410.00 |
PTCRA encodes the human pre–T cell receptor alpha (pre-TCRα), an invariant surrogate chain that pairs with TCRβ to form the pre-TCR complex during early thymocyte development. Pre-TCRα signaling coordinates β-selection, promoting thymocyte survival, proliferation, and differentiation through pathways including LCK/ZAP70-driven cascades with downstream MAPK/ERK, NF-κB, and PI3K-AKT activity. This checkpoint regulates TCRβ allelic exclusion and progression from double-negative to double-positive stages, shaping the emerging T cell repertoire. Dysregulated PTCRA expression or pre-TCR signaling has been implicated in altered lymphoid development and is frequently studied in the context of aberrant T-lineage differentiation and leukemia-associated signaling programs.
Pre-TCRα Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PTCRA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PTCRA. 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 PTCRA 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 PTCRA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.