CCDC109A Double Nickase Plasmid (h): sc-413850-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CCDC109A 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
- CCDC109A Double Nickase Plasmid (h) and CCDC109A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MCU. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CCDC109A Antibody (E-9): sc-515930
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CCDC109A Double Nickase Plasmid (h) | sc-413850-NIC | 20 µg | $410.00 | |||
CCDC109A Double Nickase Plasmid (h2) | sc-413850-NIC-2 | 20 µg | $410.00 |
MCU (CCDC109A) encodes the pore-forming subunit of the mitochondrial calcium uniporter complex that mediates Ca²⁺ uptake across the inner mitochondrial membrane. By shaping mitochondrial Ca²⁺ transients, MCU links cytosolic signaling to oxidative phosphorylation, TCA cycle dehydrogenase activation, and reactive oxygen species homeostasis, influencing ATP production and metabolic flexibility. MCU-dependent Ca²⁺ handling also interfaces with mitochondrial permeability transition and apoptosis-related pathways, affecting stress responses and cell fate decisions. Dysregulated MCU activity has been associated with altered bioenergetics and Ca²⁺ signaling in contexts including cardiometabolic dysfunction, neurodegeneration, and tumor biology, making it a frequent target in mechanistic studies of mitochondrial signaling.
CCDC109A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MCU locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MCU. 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 MCU 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 MCU-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.