LYAG Double Nickase Plasmid (h): sc-402385-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LYAG 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
- LYAG Double Nickase Plasmid (h) and LYAG Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GAA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: LYAG Antibody (G-7): sc-373745
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
LYAG Double Nickase Plasmid (h) | sc-402385-NIC | 20 µg | $410.00 | |||
LYAG Double Nickase Plasmid (h2) | sc-402385-NIC-2 | 20 µg | $410.00 |
Human GAA encodes lysosomal acid α-glucosidase (LYAG), a glycoside hydrolase required for stepwise degradation of glycogen to glucose within the lysosomal lumen. This enzyme supports lysosomal homeostasis and intersects with autophagy–lysosome pathways by preventing intralysosomal glycogen accumulation that can secondarily perturb trafficking and organelle turnover. Altered GAA activity is linked to glycogen storage pathology with prominent effects in skeletal and cardiac muscle, making the locus a key model for studying lysosomal metabolism and cellular energy balance. GAA is therefore frequently investigated in the context of lysosomal function, stress responses, and genotype–phenotype relationships in metabolic disease mechanisms.
LYAG Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GAA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GAA. 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 GAA 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 GAA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.