ITM1 Double Nickase Plasmid (h): sc-405155-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITM1 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
- ITM1 Double Nickase Plasmid (h) and ITM1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting STT3A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ITM1 Antibody (A-2): sc-390227
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ITM1 Double Nickase Plasmid (h) | sc-405155-NIC | 20 µg | $410.00 | |||
ITM1 Double Nickase Plasmid (h2) | sc-405155-NIC-2 | 20 µg | $410.00 |
Human STT3A encodes the catalytic subunit of the oligosaccharyltransferase (OST) complex responsible for co-translational N-linked glycosylation of nascent polypeptides in the endoplasmic reticulum. By transferring preassembled glycans to Asn residues within the consensus sequon, STT3A supports protein folding, quality control, and trafficking, and influences ER homeostasis and the unfolded protein response. Perturbation of STT3A-dependent glycosylation can remodel receptor maturation and signaling, alter cell-surface proteostasis, and impact secretory pathway capacity. Dysregulated N-glycosylation and ER stress pathways are implicated in diverse pathological contexts, including congenital disorders of glycosylation and cancer-associated proteostasis adaptation, making STT3A a useful target for mechanistic studies.
ITM1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the STT3A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within STT3A. 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 STT3A 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 STT3A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.