GALT Double Nickase Plasmid (h): sc-405717-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GALT 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
- GALT Double Nickase Plasmid (h) and GALT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GALT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GALT Antibody (G-1): sc-365577
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GALT Double Nickase Plasmid (h) | sc-405717-NIC | 20 µg | $410.00 | |||
GALT Double Nickase Plasmid (h2) | sc-405717-NIC-2 | 20 µg | $410.00 |
Human GALT encodes galactose-1-phosphate uridylyltransferase, a key cytosolic enzyme of the Leloir pathway that converts galactose-1-phosphate and UDP-glucose into glucose-1-phosphate and UDP-galactose. This reaction maintains galactose utilization, supports UDP-galactose pools required for glycoprotein and glycolipid biosynthesis, and helps preserve cellular metabolic homeostasis. Disruption of GALT activity leads to toxic accumulation of galactose-1-phosphate and related metabolites, linking the gene to inborn errors of carbohydrate metabolism such as classic galactosemia. As a result, GALT is widely studied in metabolic stress responses, hepatic and neuronal vulnerability, and glycoconjugate-dependent signaling and trafficking.
GALT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GALT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GALT. 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 GALT 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 GALT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.