The INPP5E Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human INPP5E gene in the HEK293T host background. This loss-of-function model enables investigation of INPP5E-dependent processes without clonal selection bias.
HEK293T cells are a derivative of HEK293 cells, stably expressing SV40 large T antigen for enhanced protein expression and viral production. Originally transformed with adenovirus type 5 DNA, these adherent epithelial cells can form primary cilia, offering a suitable system for ciliary signaling research.
INPP5E is a phosphoinositide 5-phosphatase that specifically hydrolyzes the 5-position phosphate of PIP3 and PIP2, thereby attenuating PI3K/AKT signaling by limiting PIP3-dependent AKT activation. At the primary cilium, INPP5E establishes a specialized phosphoinositide environment crucial for hedgehog pathway transduction. The protein interacts with ciliary trafficking components including ARL13B, PDE6D, and CEP164, and its expression is driven by RFX transcription factors and hedgehog pathway activity. Loss of INPP5E leads to hyperactivated AKT, mislocalization of SMO, reduced GLI transcription factor activity, and impaired expression of hedgehog target genes.
In HEK293T cells, knockout of INPP5E eliminates the key phosphoinositide regulator at the cilium, providing a well-defined genetic system to study the interplay between phosphoinositide metabolism, AKT signaling, and primary cilium assembly. The epithelial nature and ciliogenesis capacity of HEK293T cells make this model highly relevant for investigating ciliopathy mechanisms. The presence of SV40 large T antigen further enables examination of potential crosstalk between cell cycle control and PI3K/AKT pathway perturbations.
This polyclonal knockout product is suitable for a variety of research applications, including disease modeling for Joubert and MORM syndromes, mechanistic dissection of hedgehog signaling, and high-throughput screening for modulators of PI3K/AKT or ciliary function. Representative experimental approaches include western blotting for phosphorylated AKT, quantitative immunofluorescence for cilium length and localization of ARL13B, phospho-AKT ELISA, RT-qPCR quantification of GLI1 and PTCH1 transcripts, and dual-luciferase hedgehog reporter assays. For additional details, please contact Ascent Research.
