EHD3 Double Nickase Plasmid (h): sc-404685-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EHD3 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
- EHD3 Double Nickase Plasmid (h) and EHD3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EHD3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EHD3 Antibody (RR-L): sc-100723
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EHD3 Double Nickase Plasmid (h) | sc-404685-NIC | 20 µg | $410.00 | |||
EHD3 Double Nickase Plasmid (h2) | sc-404685-NIC-2 | 20 µg | $410.00 |
EHD3 (EH domain containing 3) is an ATPase of the EHD protein family that regulates endocytic recycling and membrane tubulation, coordinating trafficking between early endosomes, recycling endosomes, and the plasma membrane. Through interactions with Rab GTPase networks and membrane remodeling factors, EHD3 influences receptor turnover, cargo sorting, and signaling duration, with downstream effects on cytoskeletal organization and cell migration. Altered EHD3 expression or function has been linked to dysregulated membrane trafficking and receptor signaling in diverse disease contexts, including cancer-associated phenotypes and tissue-specific pathologies where endosomal transport is critical.
EHD3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EHD3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EHD3. 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 EHD3 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 EHD3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.