14-3-3 η Double Nickase Plasmid (h): sc-402892-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 14-3-3 η 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
- 14-3-3 η Double Nickase Plasmid (h) and 14-3-3 η Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting YWHAH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 14-3-3 η Antibody (6A12): sc-293464
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
14-3-3 η Double Nickase Plasmid (h) | sc-402892-NIC | 20 µg | $410.00 |
YWHAH encodes the human 14-3-3 η adaptor protein, a member of a conserved family that binds phosphoserine/phosphothreonine motifs to regulate the localization, stability, and activity of diverse client proteins. Through these interactions, 14-3-3 η integrates signaling nodes that control cell-cycle progression, apoptosis, stress responses, and cytoskeletal organization, with prominent connections to MAPK/ERK, PI3K–AKT, and other kinase-driven pathways. By modulating phosphorylation-dependent complexes, YWHAH contributes to neuronal development and synaptic function as well as general proteostasis and signaling fidelity. Dysregulated 14-3-3 family signaling has been linked to neuropsychiatric and neurodegenerative phenotypes and to altered growth and survival programs relevant to cancer biology, making YWHAH a useful target for mechanistic pathway studies.
14-3-3 η Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the YWHAH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within YWHAH. 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 YWHAH 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 YWHAH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.