Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
β-glucosidase Double Nickase Plasmid (h): sc-417618-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- β-glucosidase 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-glucosidase Double Nickase Plasmid (h) and β-glucosidase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GBA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: β-glucosidase Antibody (B-6): sc-166407
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
β-glucosidase Double Nickase Plasmid (h) | sc-417618-NIC | 20 µg | $410.00 | |||
β-glucosidase Double Nickase Plasmid (h2) | sc-417618-NIC-2 | 20 µg | $410.00 |
GBA encodes human β-glucosidase (glucocerebrosidase), a lysosomal hydrolase that cleaves glucosylceramide into ceramide and glucose, supporting sphingolipid turnover and membrane lipid homeostasis. This enzyme functions within the endo-lysosomal pathway and interfaces with lysosomal biogenesis and autophagy-linked processes that influence cellular proteostasis. Disruption of GBA activity leads to accumulation of glycolipid substrates and lysosomal dysfunction, a central mechanism in Gaucher disease biology. GBA variation is also widely studied for its association with synucleinopathy-related pathways, motivating mechanistic work in neuronal and myeloid model systems.
β-glucosidase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GBA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GBA. 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 GBA 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 GBA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.