Smad3 Double Nickase Plasmid (h): sc-400069-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Smad3 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
- Smad3 Double Nickase Plasmid (h) and Smad3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMAD3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Smad3 Antibody (38-Q): sc-101154
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Smad3 Double Nickase Plasmid (h) | sc-400069-NIC | 20 µg | $410.00 | |||
Smad3 Double Nickase Plasmid (h2) | sc-400069-NIC-2 | 20 µg | $410.00 |
SMAD3 encodes Smad3, a signal transducer and transcriptional regulator that mediates canonical TGF-β/Activin signaling downstream of receptor serine/threonine kinases. Upon C-terminal phosphorylation, Smad3 forms complexes with SMAD4 and translocates to the nucleus to regulate genes controlling cell-cycle arrest, extracellular matrix remodeling, differentiation, and immune modulation. Smad3 activity integrates with MAPK, PI3K/AKT, and Wnt pathways to shape context-dependent transcriptional outputs and feedback regulation. Dysregulated SMAD3 signaling has been associated with fibrosis, cancer progression, and inflammatory disease mechanisms, making it a frequent target for pathway dissection in human cell models.
Smad3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMAD3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMAD3. 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 SMAD3 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 SMAD3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.