ITM1 Double Nickase Plasmid (m): sc-421187-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITM1 Double Nickase Plasmid (m) 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 (m) and ITM1 Double Nickase Plasmid (m2) 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 (m) | sc-421187-NIC | 20 µg | $410.00 | |||
ITM1 Double Nickase Plasmid (m2) | sc-421187-NIC-2 | 20 µg | $410.00 |
Mouse Stt3a encodes a catalytic subunit of the oligosaccharyltransferase (OST) complex that mediates co-translational N-linked glycosylation of nascent polypeptides in the endoplasmic reticulum. By transferring preassembled oligosaccharides to asparagine residues within the consensus NXS/T motif, STT3A supports protein folding, quality control, and trafficking, and influences ER homeostasis and proteostasis networks. Perturbation of STT3A-dependent glycosylation can reshape the ER stress response, secretory pathway performance, and cell-surface glycoprotein composition, with downstream consequences for signaling and immune recognition. Dysregulated N-glycosylation and OST function are broadly relevant to studies of congenital disorders of glycosylation, metabolic stress, and cancer-associated remodeling of glycoproteomes, supporting mechanistic research in disease-linked pathways without implying clinical utility.
ITM1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Stt3a locus in mouse 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.