
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
nephrocystin CRISPR Activation Plasmid (h) | sc-404375-ACT | 20 µg | $397.00 |
NPHP1 encodes nephrocystin, a cilia-associated adaptor protein that localizes to the primary cilium and centrosomal/basal body compartments where it supports ciliary assembly, trafficking, and signaling competence. Nephrocystin coordinates multiprotein complexes involved in epithelial cell polarity, mechanosensation, and cytoskeletal organization, linking ciliary structure to downstream pathways that regulate differentiation and tissue homeostasis. Disrupted NPHP1 function is strongly associated with nephronophthisis-related ciliopathies and syndromic phenotypes that arise from impaired ciliary signaling and epithelial integrity. As a result, NPHP1 is frequently studied in models of renal tubulogenesis, ciliary transport dynamics, and genotype–phenotype relationships in ciliopathy biology.
nephrocystin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPHP1 expression without altering the underlying DNA sequence.
nephrocystin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPHP1 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 NPHP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous nephrocystin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPHP1 locus and enabling the study of nephrocystin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of nephrocystin pathway restoration in tumor cells with silenced or reduced NPHP1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.