Podocin CRISPR Activation Plasmid (h): sc-400930-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Podocin CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Podocin 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 Podocin CRISPR Activation Plasmid (h) and Podocin CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the NPHS2 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Podocin CRISPR Activation Plasmid (h) | sc-400930-ACT | 20 µg | $397.00 |
NPHS2 encodes podocin, an integral membrane protein enriched at the slit diaphragm of kidney podocytes where it scaffolds signaling and structural components required for glomerular filtration barrier integrity. Podocin interacts with nephrin and CD2AP within lipid raft microdomains to support actin cytoskeleton dynamics, mechanosensing, and maintenance of specialized cell junctions. Disruption of NPHS2-dependent slit diaphragm organization perturbs podocyte homeostasis and is strongly associated with proteinuric kidney disease phenotypes, making it a key node for studying filtration barrier failure. NPHS2 is therefore widely used to probe podocyte differentiation, junctional signaling pathways, and genotype–phenotype relationships in glomerular biology.
Podocin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPHS2 expression without altering the underlying DNA sequence.
Podocin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPHS2 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 NPHS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Podocin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPHS2 locus and enabling the study of Podocin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Podocin pathway restoration in tumor cells with silenced or reduced NPHS2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.