GAD-67 Double Nickase Plasmid (h): sc-400588-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GAD-67 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
- GAD-67 Double Nickase Plasmid (h) and GAD-67 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GAD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GAD-67 Antibody (F-6): sc-28376
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GAD-67 Double Nickase Plasmid (h) | sc-400588-NIC | 20 µg | $410.00 | |||
GAD-67 Double Nickase Plasmid (h2) | sc-400588-NIC-2 | 20 µg | $410.00 |
Human GAD1 encodes glutamate decarboxylase 67 (GAD-67), a key cytosolic enzyme that catalyzes the conversion of glutamate to γ-aminobutyric acid (GABA), sustaining inhibitory neurotransmission in the central nervous system. GAD-67 supports basal GABA production and influences synaptic balance, neuronal excitability, and activity-dependent plasticity through regulation of intracellular neurotransmitter pools. Altered GAD1/GAD-67 expression or regulation has been linked to disrupted excitation–inhibition homeostasis and has been studied in the context of neurodevelopmental and neuropsychiatric disorders, including schizophrenia, autism spectrum disorder, and epilepsy-related phenotypes. As a marker and functional determinant of GABAergic neuron identity, GAD-67 is commonly used to interrogate inhibitory circuit formation, maturation, and stress-responsive neurotransmitter metabolism.
GAD-67 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GAD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GAD1. 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 GAD1 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 GAD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.