Myosin Vb Double Nickase Plasmid (h): sc-403324-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Myosin Vb 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
- Myosin Vb Double Nickase Plasmid (h) and Myosin Vb Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MYO5B. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Myosin Vb Double Nickase Plasmid (h) | sc-403324-NIC | 20 µg | $410.00 | |||
Myosin Vb Double Nickase Plasmid (h2) | sc-403324-NIC-2 | 20 µg | $410.00 |
MYO5B encodes myosin Vb, an actin-based motor protein that coordinates vesicle trafficking and polarized membrane delivery through interactions with Rab GTPases and recycling endosome cargo. Myosin Vb regulates apical recycling, epithelial polarity, and transcytosis, thereby influencing tight junction organization and membrane protein localization in tissues such as intestine, liver, and kidney. Disruption of MYO5B-dependent trafficking perturbs endosomal sorting and surface expression of transporters and receptors, linking altered myosin Vb function to defects in epithelial barrier and nutrient handling. These processes place MYO5B within core cytoskeletal transport and membrane recycling pathways frequently interrogated in studies of cell polarity, endosome dynamics, and trafficking-associated disease mechanisms.
Myosin Vb Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MYO5B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MYO5B. 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 MYO5B 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 MYO5B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.