The INPP5B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human INPP5B gene in the HEK293T background. This heterogeneous pool enables robust loss-of-function studies without the artifacts of clonal isolation. Disruption of INPP5B eliminates inositol polyphosphate-5-phosphatase B activity, providing a tool for investigating phosphoinositide signaling. Supplied ready-to-use, these cells are suitable for high-throughput screening and pathway analysis.
HEK293T cells are derived from HEK293 human embryonic kidney cells transformed with adenovirus 5 DNA; they stably express the SV40 large T antigen, allowing high-level expression. Their high transfectability and rapid growth make them a standard for recombinant protein expression and viral production. The well-characterized signaling infrastructure makes HEK293T ideal for studying phosphatases like INPP5B.
INPP5B encodes an inositol polyphosphate-5-phosphatase that hydrolyzes the 5-phosphate from PI(4,5)P2 and IP3, terminating phosphoinositide signals. It acts downstream of growth factors and GPCRs, and its activity may be regulated by phosphorylation. INPP5B dephosphorylates IP3 to dampen IP3 receptor-mediated calcium release and reduces PI(4,5)P2 levels, influencing membrane phosphoinositides and adaptor protein interactions in endocytosis. Within the signaling network, it counteracts PLC, IP3 receptor, PI3K, and AKT pathways. Knockout leads to substrate accumulation, disrupting calcium homeostasis and affecting proliferation and apoptosis.
In HEK293T, INPP5B knockout creates a dysregulated phosphoinositide state relevant to Lowe syndrome and Dent disease 2, and cancer where phosphoinositide metabolism is perturbed. Loss of INPP5B can unmask oncogenic signaling via PI3K/AKT, making this model valuable for dissecting tumor suppressor roles. The high expression capacity of HEK293T facilitates biochemical and imaging studies of substrate trafficking and signaling changes, aiding drug target validation.
Applications include functional studies of phosphoinositide metabolism, calcium signaling dynamics, and cancer biology. Assays: Sanger sequencing, Western blot, IP3 ELISA, calcium flux assays with Fluo-4, immunofluorescence for PI(4,5)P2, and viability assays. These polyclonal cells offer a robust model for screening and validation. For more information, contact Ascent Research.
