The ITPRIPL2 Knockout HT29 Polyclonal Cells comprise a polyclonal population of HT29 human colorectal adenocarcinoma cells engineered via CRISPR/Cas9-mediated disruption of the ITPRIPL2 gene. This product offers a physiologically relevant model for studying the roles of ITPRIPL2 in cellular calcium dynamics and tumor progression. The polyclonal format, representing a mix of knockout alleles, avoids artifacts associated with clonal selection and enables robust statistical comparisons in functional assays. Researchers can employ this tool to dissect ITPRIPL2-dependent signaling pathways in a colorectal cancer context.
The parental HT29 cell line, derived from a primary colorectal adenocarcinoma in a 44-year-old female, is a fundamental model in colon cancer research. It displays epithelial morphology and retains key tumor features, including aberrant activation of Wnt/??-catenin and MAPK/ERK signaling. HT29 cells also exhibit defects in apoptotic machinery, making them valuable for investigating mechanisms of chemoresistance. Their extensive characterization and genetic tractability provide a solid foundation for targeted knockout experiments.
ITPRIPL2 is predicted to interact with inositol 1,4,5-trisphosphate receptors (ITPR1, ITPR2, ITPR3) and modulate endoplasmic reticulum calcium release. Within the IP3?CITPR axis, ITPRIPL2 likely binds calmodulin and FKBP12 to regulate channel activity. Downstream, calcium signals activate calmodulin, CaMKII, and NFAT transcription factors, while also triggering ER stress and caspase-3-mediated apoptosis. In HT29 cells, crosstalk with Wnt/??-catenin and MAPK/ERK pathways couples calcium flux to proliferation and survival. Knockout of ITPRIPL2 enables dissection of its integrative role in these networks.
Colorectal cancers frequently display dysregulated calcium homeostasis, contributing to apoptosis evasion. HT29 cells, which carry APC and CTNNB1 mutations, have constitutively active Wnt signaling and altered apoptosis regulators. Disrupting ITPRIPL2 in this background allows assessment of its contribution to calcium-dependent apoptotic defects and ER stress responses. The polyclonal knockout population is ideal for uncovering heterogeneous dependencies on ITPRIPL2 within cancer cell populations, potentially informing on tumor cell adaptability.
Applications include visualization of calcium fluxes via Fluo-4 AM imaging, biochemical analysis of ITPRs and phospho-proteins by Western blot, and apoptosis profiling using Annexin V/PI staining. MTT viability assays and colony formation screens evaluate drug sensitivity, while RNA-seq and RT-qPCR uncover transcriptional changes. Co-immunoprecipitation validates protein interactions, and flow cytometry provides cell cycle data. These tools support high-throughput screening for calcium modulators and chemosensitivity studies. Please contact Ascent Research for additional details or technical support.
