Rheb Double Nickase Plasmid (h): sc-400539-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rheb 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
- Rheb Double Nickase Plasmid (h) and Rheb Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RHEB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rheb Antibody (80-R): sc-130398
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rheb Double Nickase Plasmid (h) | sc-400539-NIC | 20 µg | $410.00 | |||
Rheb Double Nickase Plasmid (h2) | sc-400539-NIC-2 | 20 µg | $410.00 |
RHEB encodes Rheb, a Ras-related small GTPase that functions as a key upstream activator of mTORC1 by directly engaging the complex in response to growth factor and nutrient signaling. Through integration of cues from the TSC1/TSC2 complex and cellular energy status, Rheb helps regulate protein synthesis, autophagy, lysosomal biogenesis, and cell growth. Dysregulated RHEB–mTOR signaling has been associated with altered metabolic control and aberrant proliferation in multiple disease-relevant contexts, making Rheb a central node for studying pathway feedback and stress adaptation. In human cells, Rheb activity influences translational programs and anabolic processes that can reshape cellular phenotypes under nutrient limitation or mitogenic stimulation.
Rheb Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RHEB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RHEB. 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 RHEB 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 RHEB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.