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.
hamartin Double Nickase Plasmid (m): sc-425739-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- hamartin 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
- hamartin Double Nickase Plasmid (m) and hamartin Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Tsc1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: hamartin Antibody (C-8): sc-377386
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
hamartin Double Nickase Plasmid (m) | sc-425739-NIC | 20 µg | $410.00 | |||
hamartin Double Nickase Plasmid (m2) | sc-425739-NIC-2 | 20 µg | $410.00 |
Mouse Tsc1 encodes hamartin, a scaffold protein that forms a functional complex with TSC2 (tuberin) to act as a key negative regulator of mTORC1 signaling. Through its GAP activity toward Rheb when partnered with TSC2, the TSC1/TSC2 complex integrates inputs from PI3K–AKT, AMPK, and cellular energy or stress cues to restrain anabolic growth programs, translation, and autophagy. Loss of hamartin function disrupts homeostatic control of cell size and metabolism and is strongly linked to tuberous sclerosis complex biology, including aberrant proliferative and neurodevelopmental phenotypes in experimental models. Accordingly, Tsc1 is widely used in studies of nutrient sensing, proteostasis, synaptic function, and tumor suppressor pathways.
hamartin Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Tsc1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Tsc1. 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 Tsc1 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 Tsc1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.