GALE Double Nickase Plasmid (h): sc-408127-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GALE 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
- GALE Double Nickase Plasmid (h) and GALE Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GALE. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GALE Antibody (C-4): sc-390407
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GALE Double Nickase Plasmid (h) | sc-408127-NIC | 20 µg | $410.00 | |||
GALE Double Nickase Plasmid (h2) | sc-408127-NIC-2 | 20 µg | $410.00 |
GALE encodes UDP-galactose-4-epimerase, a cytosolic enzyme that reversibly interconverts UDP-galactose and UDP-glucose and also catalyzes the UDP-GalNAc/UDP-GlcNAc epimerization required for amino-sugar metabolism. Through these reactions, GALE supports the Leloir pathway of galactose utilization and provides nucleotide-sugar substrates for glycosylation, influencing proteoglycan and glycoprotein biosynthesis in the secretory pathway. Perturbation of GALE activity disrupts cellular carbohydrate homeostasis and can alter glycan composition, with downstream effects on protein folding, trafficking, and signaling. Human GALE dysfunction is associated with forms of galactosemia, making the gene a useful node for studying metabolic stress responses and glycosylation-linked phenotypes in model systems.
GALE Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GALE locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GALE. 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 GALE 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 GALE-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.