GP78 Double Nickase Plasmid (h): sc-402346-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GP78 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
- GP78 Double Nickase Plasmid (h) and GP78 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AMFR. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GP78 Antibody (3D9): sc-293371
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GP78 Double Nickase Plasmid (h) | sc-402346-NIC | 20 µg | $410.00 | |||
GP78 Double Nickase Plasmid (h2) | sc-402346-NIC-2 | 20 µg | $410.00 |
Human AMFR encodes GP78, an endoplasmic reticulum (ER)–resident RING-type E3 ubiquitin ligase central to ER-associated degradation (ERAD) and ubiquitin–proteasome system control of proteostasis. GP78 cooperates with E2 enzymes and ERAD components to ubiquitinate misfolded or regulated substrates, influencing unfolded protein response signaling, lipid and cholesterol homeostasis, and broader cellular stress adaptation. Through these processes, AMFR/GP78 has been studied in contexts of chronic ER stress, altered protein turnover, and pathways linked to oncogenic signaling and metastasis-associated phenotypes in model systems. Its role in ubiquitin-mediated quality control also intersects with mitochondrial dynamics and autophagy-related crosstalk, making it relevant for dissecting stress-responsive networks.
GP78 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AMFR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AMFR. 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 AMFR 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 AMFR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.