URE-B1 Double Nickase Plasmid (h): sc-404890-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- URE-B1 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
- URE-B1 Double Nickase Plasmid (h) and URE-B1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HUWE1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
URE-B1 Double Nickase Plasmid (h) | sc-404890-NIC | 20 µg | $410.00 |
HUWE1 encodes the human URE-B1 protein, a HECT-domain E3 ubiquitin ligase that catalyzes ubiquitin transfer to regulate protein stability and signaling amplitude. URE-B1 influences proteostasis and stress responses by targeting key regulators of apoptosis and DNA damage signaling, including pathways linked to p53 dynamics, cell-cycle control, and mitochondrial homeostasis. Through ubiquitin-dependent remodeling of transcriptional and repair networks, HUWE1 contributes to genome maintenance and cellular adaptation to genotoxic and proteotoxic stress. Dysregulated HUWE1 activity has been associated with altered developmental programs and oncogenic phenotypes, making it a frequently studied node in ubiquitin signaling and disease-relevant cellular remodeling.
URE-B1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HUWE1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HUWE1. 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 HUWE1 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 HUWE1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.