AGRP Double Nickase Plasmid (h): sc-402410-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AGRP 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
- AGRP Double Nickase Plasmid (h) and AGRP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AGRP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AGRP Antibody (MM0085-32B12): sc-517457
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AGRP Double Nickase Plasmid (h) | sc-402410-NIC | 20 µg | $410.00 | |||
AGRP Double Nickase Plasmid (h2) | sc-402410-NIC-2 | 20 µg | $410.00 |
AGRP encodes agouti-related protein, a neuropeptide primarily produced by hypothalamic neurons that functions as an endogenous antagonist/inverse agonist of melanocortin receptors, especially MC3R and MC4R. By opposing melanocortin signaling, AGRP helps regulate energy homeostasis, feeding behavior, and neuroendocrine responses through interconnected pathways involving POMC-derived peptides and leptin/insulin-responsive circuits. Altered AGRP signaling has been implicated in dysregulated appetite control and body-weight regulation, and it is frequently studied in the context of metabolic disease biology and hypothalamic circuitry. As a secreted peptide, AGRP also provides a tractable model for investigating neuropeptide processing, secretion, and receptor-mediated signaling in neuronal systems.
AGRP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AGRP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AGRP. 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 AGRP 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 AGRP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.