HMGCL Double Nickase Plasmid (h): sc-403841-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HMGCL 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
- HMGCL Double Nickase Plasmid (h) and HMGCL Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HMGCL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HMGCL Antibody (63Z): sc-100548
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HMGCL Double Nickase Plasmid (h) | sc-403841-NIC | 20 µg | $410.00 | |||
HMGCL Double Nickase Plasmid (h2) | sc-403841-NIC-2 | 20 µg | $410.00 |
HMGCL encodes 3-hydroxymethyl-3-methylglutaryl-CoA lyase, a mitochondrial enzyme that catalyzes the terminal step of ketogenesis and leucine catabolism by converting HMG-CoA to acetoacetate and acetyl-CoA. This activity links mitochondrial acetyl-CoA handling to cellular energy balance during fasting and other metabolic stresses, and interfaces with broader pathways controlling fatty acid oxidation, anaplerosis, and redox homeostasis. Altered HMGCL function is associated with inborn errors of metabolism affecting ketone body production and branched-chain amino acid degradation, making it relevant to studies of metabolic adaptation and mitochondrial physiology. In cancer biology and immunometabolism, HMGCL-dependent ketone metabolism has been explored as a contributor to nutrient utilization and signaling states under hypoxia or nutrient limitation.
HMGCL Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HMGCL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HMGCL. 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 HMGCL 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 HMGCL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.