ITPRIP Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ITPRIP gene has been disrupted via CRISPR/Cas9-mediated gene targeting. This product provides a heterogeneous pool of A-549 cells carrying diverse edits at the ITPRIP locus, enabling functional loss-of-function studies without clonal selection. The polyclonal format captures the variability of the knockout population, making it suitable for applications where pooled genetic perturbation is desired, such as high-throughput screens or pathway-level analyses. The product is supplied as a ready-to-use polyclonal culture, prepared under controlled conditions to maintain viability and proliferative capacity.
Host A-549 cells originate from human lung adenocarcinoma tissue of a 58-year-old Caucasian male and exhibit an epithelial morphology reminiscent of alveolar Type II pneumocytes. This adherent cell line is widely employed as a model system for lung adenocarcinoma research, including investigations into oncogenic signaling, apoptosis regulation, and chemotherapeutic drug resistance. The cells retain key features of transformed lung epithelium, such as expression of surfactant proteins and active proliferation, making them a robust platform for studying tumor cell biology and therapeutic interventions.
The ITPRIP gene product is a key modulator of intracellular calcium dynamics and autophagy. ITPRIP binds directly to inositol 1,4,5-trisphosphate receptors (IP3R isoforms ITPR1, ITPR2, ITPR3), sensitizing these channels to low concentrations of IP3 and thereby potentiating calcium release from the endoplasmic reticulum. Independent of its calcium-regulating role, ITPRIP interacts with the autophagy-related protein MAP1LC3C (LC3C) to promote autophagosome formation and, through association with RPTOR and ULK1, inhibits mTORC1 activity. This dual functionality places ITPRIP at the intersection of calcium signaling and nutrient-sensing pathways, with downstream consequences for transcription factors such as NFAT and CREB, as well as for the core autophagy machinery, including BECN1 and ATG5-ATG12.
In the A-549 lung adenocarcinoma context, disruption of ITPRIP unlinks two critical homeostatic networks. Loss of ITPRIP attenuates IP3-dependent calcium oscillations and LC3C-mediated autophagy, thereby altering cellular responses to metabolic stress, growth factors, and chemotherapeutics. This knockout model permits the dissection of ITPRIP-dependent contributions to oncogenic calcium signaling and to the cytoprotective autophagy often observed in drug-resistant lung cancer cells. Consequently, the ITPRIP Knockout A-549 Polyclonal Cells provide a physiologically relevant backdrop for examining how calcium-mTORC1-autophagy crosstalk influences tumor progression and treatment susceptibility.
Research applications span multiple disciplines. These cells enable calcium imaging studies with Fluo-4 to quantify ER Ca2+ release, co-immunoprecipitation experiments to map ITPRIP-IP3R or ITPRIP-LC3C interactions, and autophagy flux assays using bafilomycin A1 to assess autophagic turnover. Additional uses include mTORC1 activity measurements via phospho-S6K immunoblotting, high-throughput screening for autophagy modulators, and mechanistic investigations into drug resistance in lung adenocarcinoma. Representative assays combine Western blotting for LC3B/LC3C lipidation, RT-qPCR verification of ITPRIP disruption, and immunofluorescence detection of autophagosome markers. These polyclonal knockout cells serve as a versatile tool for elucidating ITPRIP-related biology. For further information, contact Ascent Research.
