mTOR Double Nickase Plasmid (h): sc-400140-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mTOR 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
- mTOR Double Nickase Plasmid (h) and mTOR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MTOR. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: p-mTOR Antibody (59.Ser 2448): sc-293133
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mTOR Double Nickase Plasmid (h) | sc-400140-NIC | 20 µg | $410.00 | |||
mTOR Double Nickase Plasmid (h2) | sc-400140-NIC-2 | 20 µg | $410.00 |
MTOR encodes the serine/threonine kinase mTOR, a central integrator of growth factor, nutrient, energy, and stress cues that coordinates cellular metabolism and homeostasis. mTOR functions within mTORC1 and mTORC2 to regulate protein synthesis and cell growth via S6K and 4E-BP1, autophagy through ULK1 signaling, and cytoskeletal organization and survival signaling through AKT and SGK pathways. Dysregulated mTOR signaling is implicated in altered proliferation and metabolic reprogramming across cancer biology, and contributes to neurodevelopmental and neurodegenerative phenotypes as well as cardiometabolic dysfunction. Because mTOR sits at the nexus of PI3K–AKT–mTOR and AMPK signaling, MTOR perturbation is widely used to interrogate pathway crosstalk, stress responses, and translational control.
mTOR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MTOR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MTOR. 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 MTOR 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 MTOR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.