Description

The NPHP3 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line for loss-of-function studies of NPHP3. This product provides a genetically stable model to investigate NPHP3 function in ciliary signaling and renal developmental pathways. Engineered through CRISPR/Cas9-mediated gene disruption, it avoids the variability of transient suppression approaches and enables consistent experimental outcomes. The cell line offers a valuable tool for research into ciliopathies such as nephronophthisis type 3 and associated syndromes.

The parental HEK293T cell line originates from human embryonic kidney 293 cells and constitutively expresses the SV40 large T antigen. This feature permits high-level, transient protein expression and supports episomal replication of plasmids carrying the SV40 ori, making it a workhorse for viral vector production. HEK293T cells maintain an epithelial phenotype and primary cilia, providing a relevant cellular context for kidney biology studies and facilitating the analysis of cilia-dependent processes.

NPHP3 localizes to the ciliary transition zone, where it interacts with NPHP1, NPHP4, INVS, and RPGRIP1L to form a barrier that controls signaling molecule entry and exit. Its expression is regulated by RFX transcription factors and HNF1B. NPHP3 regulates downstream signaling, including the Wnt/??-catenin pathway through components such as WNT5A, FZD, and DVL, and the planar cell polarity pathway by modulating RhoA and JNK activity. Gene disruption leads to altered transcriptional output, exemplified by changes in AXIN2 expression, and impairs ciliary gating of signaling mediators.

In HEK293T cells, NPHP3 knockout recapitulates key molecular features of ciliopathy-related signaling defects. The renal epithelial origin of these cells makes the knockout line particularly suitable for modeling nephronophthisis type 3 and related renal developmental disorders. The presence of primary cilia in HEK293T allows direct investigation of ciliary assembly, protein trafficking, and downstream signaling events disrupted upon loss of NPHP3. This system enables high-resolution studies of ciliary dysfunction within a disease-relevant cellular environment.

This cell line supports diverse applications including immunofluorescence analysis of ciliary protein localization, western blotting for phospho-JNK as a planar cell polarity readout, and RT-qPCR to quantify AXIN2 expression indicative of Wnt pathway activity. It is also suited for ciliogenesis assays, cell migration studies, and drug sensitivity screening focused on Wnt pathway inhibitors. Together, these applications facilitate mechanistic dissection of ciliopathies and preclinical evaluation of therapeutic strategies. For further details, contact Ascent Research.