KGA Double Nickase Plasmid (h): sc-402575-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- KGA 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
- KGA Double Nickase Plasmid (h) and KGA Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GLS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: KGA Antibody (IJ-2): sc-100533
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
KGA Double Nickase Plasmid (h) | sc-402575-NIC | 20 µg | $410.00 | |||
KGA Double Nickase Plasmid (h2) | sc-402575-NIC-2 | 20 µg | $410.00 |
Human GLS encodes kidney-type glutaminase (KGA), a mitochondrial enzyme that hydrolyzes glutamine to glutamate, supplying carbon and nitrogen for the TCA cycle, nucleotide biosynthesis, and redox balance via glutathione production. KGA activity connects glutamine metabolism to cellular bioenergetics, anaplerosis, and signaling networks that regulate proliferation and stress adaptation. Altered GLS/KGA function has been associated with metabolic reprogramming observed in cancer models and with susceptibility in neurological and immune-related phenotypes linked to glutamate handling. Studying GLS supports mechanistic work on nutrient utilization, mitochondrial metabolism, and pathway crosstalk affecting oxidative stress and cell fate decisions.
KGA Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GLS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GLS. 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 GLS 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 GLS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.