MIB1 Double Nickase Plasmid (h): sc-404769-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MIB1 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
- MIB1 Double Nickase Plasmid (h) and MIB1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MIB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MIB1 Antibody (D-6): sc-393551
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MIB1 Double Nickase Plasmid (h) | sc-404769-NIC | 20 µg | $410.00 | |||
MIB1 Double Nickase Plasmid (h2) | sc-404769-NIC-2 | 20 µg | $410.00 |
MIB1 encodes mindbomb E3 ubiquitin protein ligase 1, a RING-type E3 ubiquitin ligase that ubiquitinates Notch ligands such as DLL1 and JAG1 to promote ligand endocytosis and productive Notch receptor activation. Through regulation of Notch signaling amplitude and timing, MIB1 influences cell fate decisions, differentiation, and tissue patterning, with downstream effects on transcriptional programs controlling proliferation and lineage commitment. MIB1 also intersects with ubiquitin–proteasome homeostasis and endocytic trafficking pathways that shape receptor–ligand dynamics at the cell surface. Genetic and functional perturbation of MIB1 has been associated with altered developmental signaling and disease-relevant phenotypes, including contexts where Notch pathway dysregulation contributes to oncogenic and cardiovascular biology.
MIB1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MIB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MIB1. 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 MIB1 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 MIB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.