NET-4 Double Nickase Plasmid (h): sc-404621-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NET-4 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
- NET-4 Double Nickase Plasmid (h) and NET-4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TSPAN5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NET-4 Double Nickase Plasmid (h) | sc-404621-NIC | 20 µg | $410.00 | |||
NET-4 Double Nickase Plasmid (h2) | sc-404621-NIC-2 | 20 µg | $410.00 |
TSPAN5 encodes tetraspanin-5 (NET-4), a four-pass membrane protein that organizes tetraspanin-enriched microdomains to coordinate receptor clustering, cell–cell communication, and membrane trafficking. NET-4 has been implicated in regulating cell adhesion and migration through its interactions with partner proteins at the plasma membrane, influencing signaling processes linked to cytoskeletal remodeling and vesicular transport. In human tissues, TSPAN5 expression has been associated with neuronal development and synaptic organization, consistent with broader roles for tetraspanins in modulating receptor dynamics and intercellular signaling. Dysregulated tetraspanin networks, including TSPAN5-related microdomain changes, are studied in the context of altered differentiation programs and disease-relevant signaling states.
NET-4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TSPAN5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TSPAN5. 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 TSPAN5 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 TSPAN5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.