β-Gal Double Nickase Plasmid (h): sc-401194-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- β-Gal 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
- β-Gal Double Nickase Plasmid (h) and β-Gal Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GLB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: β-Gal Antibody (B-12): sc-377257
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
β-Gal Double Nickase Plasmid (h) | sc-401194-NIC | 20 µg | $410.00 | |||
β-Gal Double Nickase Plasmid (h2) | sc-401194-NIC-2 | 20 µg | $410.00 |
GLB1 encodes human β-galactosidase (β-Gal), a lysosomal exoglycosidase that hydrolyzes terminal β-galactose residues from GM1 ganglioside, glycoproteins, and glycosaminoglycans. This activity supports lysosomal catabolic flux and glycosphingolipid turnover, intersecting with endo-lysosomal trafficking, autophagy-lysosome function, and cellular quality-control pathways. Disruption of GLB1 perturbs substrate clearance and is associated with lysosomal storage disorders including GM1 gangliosidosis and Morquio B disease, making it a useful model gene for studying lysosomal dysfunction. GLB1 is therefore frequently investigated in the context of neurodegeneration-linked stress responses, metabolic remodeling, and the cellular consequences of impaired glycan processing.
β-Gal Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GLB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GLB1. 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 GLB1 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 GLB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.