Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
MCAD Double Nickase Plasmid (h): sc-402708-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MCAD 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
- MCAD Double Nickase Plasmid (h) and MCAD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACADM. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MCAD Antibody (E-8): sc-271931
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MCAD Double Nickase Plasmid (h) | sc-402708-NIC | 20 µg | $410.00 | |||
MCAD Double Nickase Plasmid (h2) | sc-402708-NIC-2 | 20 µg | $410.00 |
ACADM encodes medium-chain acyl-CoA dehydrogenase (MCAD), a mitochondrial flavoprotein that catalyzes the first dehydrogenation step of medium-chain fatty acyl-CoAs during β-oxidation. MCAD activity supports energy production from fatty acids, contributing to metabolic flexibility during fasting and other high-demand states, and interfaces with broader mitochondrial redox balance and acetyl-CoA supply. Disruption of ACADM is linked to impaired fatty acid oxidation and accumulation of medium-chain acylcarnitines, a biochemical signature widely used in metabolic research. ACADM/MCAD is therefore a key node for studying mitochondrial metabolism, nutrient sensing, and lipid-derived signaling in human cell models.
MCAD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACADM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACADM. 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 ACADM 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 ACADM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.