Smad4 Double Nickase Plasmid (h): sc-400110-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Smad4 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
- Smad4 Double Nickase Plasmid (h) and Smad4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMAD4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Smad4 Antibody (B-8): sc-7966
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Smad4 Double Nickase Plasmid (h) | sc-400110-NIC | 20 µg | $410.00 | |||
Smad4 Double Nickase Plasmid (h2) | sc-400110-NIC-2 | 20 µg | $410.00 |
SMAD4 encodes Smad4, a central co-mediator of canonical TGF-β and BMP signaling that forms heteromeric complexes with receptor-regulated SMADs to control transcriptional programs governing proliferation, differentiation, and epithelial–mesenchymal dynamics. Upon ligand-induced receptor activation, Smad4-containing complexes accumulate in the nucleus and integrate inputs from pathways such as MAPK, WNT/β-catenin, and PI3K/AKT to shape context-dependent gene expression. Disruption of SMAD4 alters TGF-β–dependent growth control and tissue homeostasis and is frequently linked to tumor suppressor dysfunction in gastrointestinal and pancreatic malignancies. As a signaling hub, SMAD4 is widely studied for its roles in development, fibrosis-related transcriptional states, and immune-modulatory gene regulation.
Smad4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMAD4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMAD4. 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 SMAD4 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 SMAD4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.