HYPE Double Nickase Plasmid (h): sc-406626-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HYPE 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
- HYPE Double Nickase Plasmid (h) and HYPE Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FICD. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HYPE Antibody (G-7): sc-515368
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HYPE Double Nickase Plasmid (h) | sc-406626-NIC | 20 µg | $410.00 | |||
HYPE Double Nickase Plasmid (h2) | sc-406626-NIC-2 | 20 µg | $410.00 |
FICD encodes HYPE, an endoplasmic reticulum–associated Fic domain enzyme that catalyzes AMPylation and de-AMPylation of target proteins, most prominently the HSPA5/GRP78 chaperone. By modulating GRP78 activity, HYPE influences proteostasis, unfolded protein response signaling, and ER stress adaptation, integrating with pathways that coordinate protein folding and quality control. Dysregulated ER homeostasis and stress signaling linked to FICD/HYPE activity has been associated with cellular vulnerability in contexts such as neurodegeneration, metabolic dysfunction, and cancer biology, making it relevant for mechanistic studies of stress-responsive signaling networks.
HYPE Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FICD locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FICD. 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 FICD 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 FICD-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.