TMEM65 Double Nickase Plasmid (h): sc-406584-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TMEM65 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
- TMEM65 Double Nickase Plasmid (h) and TMEM65 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TMEM65. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TMEM65 Double Nickase Plasmid (h) | sc-406584-NIC | 20 µg | $410.00 | |||
TMEM65 Double Nickase Plasmid (h2) | sc-406584-NIC-2 | 20 µg | $410.00 |
TMEM65 encodes a mitochondrial inner membrane protein implicated in maintaining mitochondrial architecture and supporting oxidative phosphorylation, with downstream effects on ATP production and cellular redox balance. By influencing mitochondrial protein organization and respiratory chain function, TMEM65 contributes to processes such as membrane potential regulation and mitochondrial homeostasis. Altered TMEM65 activity has been associated with bioenergetic dysfunction and phenotypes consistent with mitochondrial disease mechanisms, making it relevant for studies of neuromuscular and cardiometabolic stress responses. Its mitochondrial localization also positions it as a useful node for investigating signaling links between mitochondrial dysfunction, proteostasis, and cellular adaptation pathways.
TMEM65 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TMEM65 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TMEM65. 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 TMEM65 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 TMEM65-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.