CRF Double Nickase Plasmid (h): sc-400770-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CRF 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
- CRF Double Nickase Plasmid (h) and CRF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CRH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CRF Antibody (2B11): sc-293187
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CRF Double Nickase Plasmid (h) | sc-400770-NIC | 20 µg | $410.00 | |||
CRF Double Nickase Plasmid (h2) | sc-400770-NIC-2 | 20 µg | $410.00 |
Human CRH encodes corticotropin-releasing hormone (CRF), a key neuropeptide that initiates the hypothalamic–pituitary–adrenal (HPA) axis response to physiological and psychological stress. CRF signaling through CRHR1/CRHR2 activates cAMP/PKA and downstream CREB-regulated transcription, shaping endocrine output, autonomic function, and behavioral adaptations. Beyond the central nervous system, CRH/CRF can modulate immune activity and inflammatory signaling in peripheral tissues. Dysregulated CRH–CRF pathways are implicated in stress-related neuropsychiatric phenotypes and altered neuroendocrine homeostasis, and are studied in contexts including mood and anxiety circuitry, sleep regulation, and metabolic adaptation.
CRF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CRH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CRH. 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 CRH 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 CRH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.