CD133 Double Nickase Plasmid (m): sc-437381-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD133 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
- CD133 Double Nickase Plasmid (m) and CD133 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Prom1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CD133 Antibody (E-11): sc-365537
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD133 Double Nickase Plasmid (m) | sc-437381-NIC | 20 µg | $410.00 | |||
CD133 Double Nickase Plasmid (m2) | sc-437381-NIC-2 | 20 µg | $410.00 |
Prom1 encodes the pentaspan membrane glycoprotein CD133, a cholesterol-binding protein enriched in plasma membrane protrusions that helps organize membrane microdomains and regulate cell polarity. In mouse tissues, CD133 is widely used as a marker of stem and progenitor compartments and is linked to programs controlling self-renewal, differentiation, and tissue regeneration. CD133-associated signaling intersects with pathways governing epithelial organization and cytoskeletal dynamics, and altered Prom1 expression is reported in models of retinal degeneration, neurodevelopmental defects, and tumor-initiating cell phenotypes. Functional studies of Prom1 therefore inform mechanisms of lineage commitment, barrier-forming epithelia, and stem-like cellular states relevant to disease biology.
CD133 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Prom1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Prom1. 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 Prom1 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 Prom1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.