TMEM66 Double Nickase Plasmid (h): sc-412236-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TMEM66 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
- TMEM66 Double Nickase Plasmid (h) and TMEM66 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SARAF. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TMEM66 Double Nickase Plasmid (h) | sc-412236-NIC | 20 µg | $410.00 |
SARAF (SOCE-associated regulatory factor), also known as TMEM66, is an endoplasmic reticulum membrane protein that modulates store-operated calcium entry by interacting with STIM1 and contributing to Ca2+-dependent inactivation of Orai1 channels. By tuning cytosolic calcium signaling, SARAF influences downstream pathways linked to gene expression, secretion, metabolism, and stress adaptation, including ER homeostasis and unfolded protein response dynamics. Dysregulated SOCE and ER calcium handling are implicated in multiple disease-relevant processes such as aberrant proliferation, altered immune cell activation, and vulnerability to oxidative and proteotoxic stress. SARAF/TMEM66 is therefore a useful target for mechanistic studies connecting calcium flux to cell fate decisions and signaling network rewiring.
TMEM66 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SARAF locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SARAF. 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 SARAF 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 SARAF-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.