Smad5 Double Nickase Plasmid (h): sc-400443-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Smad5 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
- Smad5 Double Nickase Plasmid (h) and Smad5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMAD5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Smad5 Antibody (YY-6): sc-101151
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Smad5 Double Nickase Plasmid (h) | sc-400443-NIC | 20 µg | $410.00 | |||
Smad5 Double Nickase Plasmid (h2) | sc-400443-NIC-2 | 20 µg | $410.00 |
SMAD5 encodes the receptor-regulated transcription factor Smad5, a central mediator of BMP signaling downstream of type I/II serine–threonine kinase receptors. Upon BMP-dependent phosphorylation, Smad5 forms complexes with SMAD4 and translocates to the nucleus to control gene programs that regulate proliferation, differentiation, apoptosis, and lineage specification. SMAD5 activity is particularly relevant to hematopoietic and endothelial biology and to developmental patterning processes controlled by BMP gradients. Dysregulated SMAD5/BMP signaling has been associated with congenital anomalies and cancer-related pathway remodeling, making SMAD5 a useful node for dissecting context-specific transcriptional outputs.
Smad5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMAD5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMAD5. 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 SMAD5 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 SMAD5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.