Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
UBC9 Double Nickase Plasmid (h): sc-400859-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- UBC9 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
- UBC9 Double Nickase Plasmid (h) and UBC9 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting UBE2I. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: UBC9 Antibody (C-12): sc-271057
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
UBC9 Double Nickase Plasmid (h) | sc-400859-NIC | 20 µg | $410.00 | |||
UBC9 Double Nickase Plasmid (h2) | sc-400859-NIC-2 | 20 µg | $410.00 |
UBE2I encodes UBC9, the sole E2 SUMO-conjugating enzyme that transfers SUMO to target proteins to regulate protein stability, subcellular localization, and macromolecular interactions. Through SUMOylation, UBC9 influences DNA damage signaling and repair, cell-cycle progression, chromatin organization, transcriptional programs, and nucleocytoplasmic transport, integrating stress-responsive pathways with proteostasis. Perturbation of UBC9-dependent SUMOylation has been linked to dysregulated genome maintenance and altered transcriptional control, and aberrant SUMO pathway activity is frequently observed in cancer biology and neurodegeneration-associated cellular stress phenotypes. As a central node in the SUMO cascade, UBC9 serves as a useful entry point for dissecting SUMO-dependent regulation of key substrates and pathway crosstalk.
UBC9 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the UBE2I locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within UBE2I. 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 UBE2I 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 UBE2I-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.