The ITPR1 Knockout HCT 116 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma line, with targeted disruption of the ITPR1 gene. This polyclonal pool provides a heterogeneous loss-of-function model for investigating ITPR1-dependent processes, avoiding clonal selection bias.
The host HCT 116 cell line is a well-established colorectal carcinoma model, harboring a KRAS G13D mutation and microsatellite instability (MSI). These features drive constitutive MAPK signaling and a hypermutable phenotype, making the line ideal for studying tumorigenesis, intracellular signaling, and drug responses.
ITPR1 encodes the inositol 1,4,5-trisphosphate receptor (IP3R1), an endoplasmic reticulum Ca2? release channel. Stimuli from GPCRs and RTKs activate phospholipase C to generate IP3, which gates ITPR1, triggering Ca2? efflux. This release is modulated by PKA and PKC, and signals through calmodulin, calcineurin, CAMKII, and the mitochondrial calcium uniporter, affecting transcription factors NFAT and CREB, and caspases. Interacting partners include Bcl-2, Beclin-1, FKBP12, and IRBIT, linking ITPR1 to apoptosis, autophagy, and survival pathways.
In the KRAS-mutant, MSI HCT 116 context, ITPR1 knockout enables dissection of calcium signaling’s role in colorectal cancer. ITPR1-mediated Ca2? fluxes influence KRAS-driven proliferation, apoptotic resistance, and autophagy, processes that affect tumor progression and chemosensitivity. This model is particularly relevant for exploring how altered ER-mitochondria Ca2? transfer contributes to drug resistance in MSI colorectal tumors.
Applications include calcium imaging with Fluo-4 or Fura-2, Western blot and RT-qPCR validation of knockout, flow cytometry for apoptosis (Annexin V/PI), and autophagy flux assays. Cell viability assays support drug sensitivity screening (e.g., 5-fluorouracil, oxaliplatin), while co-immunoprecipitation facilitates protein interaction studies. The model also aids in spinocerebellar ataxia research. For further information, contact Ascent Research.
