The ITPR3 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9 gene-edited polyclonal cell population derived from the human A-549 lung adenocarcinoma epithelial line, engineered to disrupt expression of the inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) gene. As a polyclonal knockout product, this population comprises a heterogeneous mixture of cells carrying distinct CRISPR/Cas9-induced mutations that abrogate functional ITPR3 protein, enabling phenotype analysis in a context that mitigates clonal selection bias while maintaining a native-like cellular background.
The A-549 host cell line was originally isolated from a 58-year-old Caucasian male with lung adenocarcinoma and grows as an adherent epithelial monolayer. Extensively employed in respiratory disease modeling, drug metabolism investigations, and oncogenic signaling studies, A-549 cells provide a robust and well-characterized platform for generating gene knockouts to dissect molecular pathways underlying non-small cell lung cancer biology.
ITPR3 encodes the type 3 inositol 1,4,5-trisphosphate receptor, an endoplasmic reticulum-resident calcium-release channel. Following activation of phospholipase C (PLC) downstream of G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs) such as EGF and PDGF, the second messenger IP3 binds to ITPR3, triggering calcium mobilization into the cytosol. This calcium signal activates critical effectors including calmodulin, calcineurin, and Ca2+/calmodulin-dependent protein kinase II (CaMKII), which in turn regulate transcription factors such as NFAT, NF-??B, and CREB. ITPR3 is modulated by interacting partners like FKBP12, Homer, and Bcl-2, and integrates inputs from PKA and PKC, thereby coupling calcium dynamics to pathways including NF-??B, MAPK, and mTOR that govern cell proliferation, apoptosis, and motility.
In the A-549 lung adenocarcinoma context, ITPR3-mediated calcium signaling is closely tied to tumorigenic processes. Loss of ITPR3 disrupts calcium-dependent activation of downstream cascades such as NF-??B and MAPK, potentially impairing cell survival, migration, and invasive capacity. Consequently, this knockout model offers a physiologically relevant system to investigate the contribution of IP3 receptor-driven calcium flux to lung cancer progression, metastasis, and sensitivity to therapeutics.
This polyclonal knockout product is well-suited for a range of experimental applications, including calcium imaging with Fluo-4, cell viability (MTT) and apoptosis (Annexin V) assays, and transwell migration studies. It is also compatible with western blotting, RT?qPCR, immunofluorescence, and co?immunoprecipitation for validating engagement of downstream targets and pathways. These tools collectively enable in-depth exploration of ITPR3 functions in drug resistance, cancer metastasis, and oncogenic signaling. For further product details, including lot-specific knockout efficiency and validation data, please contact Ascent Research.
