BCKDE1A Double Nickase Plasmid (h): sc-417950-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BCKDE1A 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
- BCKDE1A Double Nickase Plasmid (h) and BCKDE1A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BCKDHA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: BCKDE1A Antibody (H-5): sc-271538
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BCKDE1A Double Nickase Plasmid (h) | sc-417950-NIC | 20 µg | $410.00 | |||
BCKDE1A Double Nickase Plasmid (h2) | sc-417950-NIC-2 | 20 µg | $410.00 |
BCKDHA encodes the E1α subunit of the branched-chain α-ketoacid dehydrogenase (BCKDH) complex, a mitochondrial enzyme system that catalyzes the oxidative decarboxylation of branched-chain amino acid–derived α-ketoacids. This activity is central to valine, leucine, and isoleucine catabolism and links amino acid turnover to acetyl-CoA and succinyl-CoA production, influencing mitochondrial redox balance and energy metabolism. BCKDHA function is regulated through BCKDH complex phosphorylation dynamics and mitochondrial cofactor availability, integrating nutrient sensing with metabolic flux. Disruption of BCKDHA is associated with impaired branched-chain amino acid metabolism and inborn errors of metabolism such as maple syrup urine disease, making it relevant for studies of metabolic stress, mitochondrial dysfunction, and amino acid–driven signaling.
BCKDE1A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BCKDHA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BCKDHA. 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 BCKDHA 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 BCKDHA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.