H-Ras Double Nickase Plasmid (h): sc-400204-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- H-Ras 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
- H-Ras Double Nickase Plasmid (h) and H-Ras Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HRAS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: H-Ras Antibody (M90): sc-53959
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
H-Ras Double Nickase Plasmid (h) | sc-400204-NIC | 20 µg | $410.00 | |||
H-Ras Double Nickase Plasmid (h2) | sc-400204-NIC-2 | 20 µg | $410.00 |
HRAS encodes the human small GTPase H-Ras, a membrane-associated molecular switch that cycles between GDP- and GTP-bound states to transmit signals from receptor tyrosine kinases and other upstream inputs. Activated H-Ras engages RAF–MEK–ERK and PI3K–AKT signaling, integrating cues that regulate proliferation, differentiation, cytoskeletal dynamics, and survival. Tight control of GTP hydrolysis by GEFs and GAPs is essential for normal signaling amplitude and duration, and dysregulation of RAS signaling is strongly linked to oncogenic transformation and aberrant growth programs. HRAS is therefore widely used as a model locus for dissecting MAPK pathway regulation, feedback control, and context-dependent signal output in human cells.
H-Ras Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HRAS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HRAS. 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 HRAS 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 HRAS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.