Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Smad8 CRISPR Activation Plasmid (h): sc-402119-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Smad8 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Smad8 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Smad8 CRISPR Activation Plasmid (h) and Smad8 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the SMAD9 transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Smad8 Antibody (3E5): sc-293413
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Smad8 CRISPR Activation Plasmid (h) | sc-402119-ACT | 20 µg | $397.00 |
SMAD9 encodes Smad8, a receptor-regulated SMAD that transduces bone morphogenetic protein (BMP) signals downstream of type I/II serine/threonine kinase receptors. Following BMP receptor activation, Smad8 is phosphorylated, forms complexes with SMAD4, and accumulates in the nucleus to modulate transcriptional programs controlling cell fate specification, differentiation, and tissue homeostasis. SMAD9-regulated BMP/TGF-β pathway outputs intersect with developmental patterning and osteogenic gene networks, making SMAD9 a frequent target in studies of skeletal biology and lineage commitment. Dysregulated BMP–SMAD signaling involving SMAD9 has been associated with human developmental and cardiopulmonary phenotypes and is broadly relevant to research on fibrosis, inflammation, and tumor microenvironment signaling without implying clinical outcomes.
Smad8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMAD9 expression without altering the underlying DNA sequence.
Smad8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMAD9 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the SMAD9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Smad8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMAD9 locus and enabling the study of Smad8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Smad8 pathway restoration in tumor cells with silenced or reduced SMAD9 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.