GP78 Double Nickase Plasmid (m): sc-423842-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GP78 Double Nickase Plasmid (m) 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 (m) and GP78 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Amfr. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GP78 Double Nickase Plasmid (m) | sc-423842-NIC | 20 µg | $410.00 | |||
GP78 Double Nickase Plasmid (m2) | sc-423842-NIC-2 | 20 µg | $410.00 |
Amfr encodes GP78, an endoplasmic reticulum (ER)-resident E3 ubiquitin ligase that functions as a key component of ER-associated degradation (ERAD) by ubiquitinating misfolded or surplus proteins for proteasomal turnover. GP78 cooperates with E2 enzymes and factors such as p97/VCP to coordinate retrotranslocation and clearance of ER client proteins, thereby supporting proteostasis and limiting ER stress signaling. Through its roles in ubiquitin-dependent quality control, GP78 influences pathways linked to lipid metabolism, mitochondrial homeostasis, and stress-adaptive responses. Dysregulated ERAD and ubiquitin signaling involving GP78 have been associated with disease-relevant processes including metabolic imbalance and proteotoxic stress that are frequently studied in neurodegeneration and cancer biology.
GP78 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Amfr locus in mouse 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.