Giantin Double Nickase Plasmid (h): sc-418145-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Giantin 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
- Giantin Double Nickase Plasmid (h) and Giantin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GOLGB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Giantin Antibody (2388C3a): sc-81279
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Giantin Double Nickase Plasmid (h) | sc-418145-NIC | 20 µg | $410.00 | |||
Giantin Double Nickase Plasmid (h2) | sc-418145-NIC-2 | 20 µg | $410.00 |
GOLGB1 encodes Giantin, a large coiled-coil golgin that anchors to Golgi membranes and helps organize Golgi architecture by capturing transport vesicles and coordinating tethering events. Giantin supports ER-to-Golgi and intra-Golgi trafficking, contributes to glycosylation and secretory pathway homeostasis, and influences the spatial organization of Golgi enzymes required for protein processing. Disruption of golgin function, including Giantin-dependent Golgi maintenance, is linked to altered extracellular matrix secretion, stress responses in the secretory pathway, and phenotypes relevant to ciliogenesis and tissue morphogenesis. As a structural and trafficking regulator, Giantin is widely used in cell biology studies of Golgi integrity, vesicle dynamics, and protein maturation in human cells.
Giantin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GOLGB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GOLGB1. 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 GOLGB1 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 GOLGB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.