GRINA Knockout HeLa Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population in which the GRINA gene has been disrupted, providing a loss-of-function model for investigating apoptosis and endoplasmic reticulum (ER) stress pathways. This polyclonal cell population is derived from the HeLa host cell line and enables functional studies of GRINA-dependent signaling without the need for single-cell clone isolation. The knockout model is generated through CRISPR/Cas9-mediated gene disruption, ensuring targeted abrogation of GRINA expression across the cell pool. Researchers can use this product to analyze GRINA??s role in cell survival, calcium homeostasis, and the unfolded protein response in a reproducible human cancer cell context.
The host cell line, HeLa, is a widely used human cervical adenocarcinoma cell line that contains human papillomavirus type 18 (HPV-18) sequences. As an epithelial cell model, HeLa cells exhibit robust proliferation and are permissive to genetic manipulation, making them a workhorse for cancer biology, virology, and cellular signaling studies. The presence of HPV-18 oncoproteins E6 and E7 disrupts p53 and retinoblastoma protein functions, creating a unique background for apoptosis and cell cycle investigations. This immortalized cell line thus provides a relevant platform for examining how GRINA modulates cell death mechanisms in the context of viral oncogenesis and cervical cancer.
GRINA (also known as TMBIM3) encodes a transmembrane BAX inhibitor motif-containing protein that localizes to both the ER and mitochondria. It functions as a key inhibitor of apoptosis by directly interacting with BAX and BAK, blocking their oligomerization and subsequent cytochrome c release from mitochondria. Additionally, GRINA regulates ER calcium homeostasis through interactions with inositol 1,4,5-trisphosphate (IP3) receptors and ryanodine receptors, thereby influencing the unfolded protein response. Upstream, GRINA is regulated by ER stress sensors ATF6, IRE1, and PERK, as well as by calcium influx, oxidative stress, and the transcription factor CREB1. Downstream, its activity suppresses caspase activation and cytochrome c release, with molecular interactions involving BCL-2 family members, presenilin 1, and the chaperone GRP78/BiP. Key pathway components include CHOP, caspase-3, and the unfolded protein response mediators.
In the HeLa cervical cancer context, GRINA knockout offers a powerful system to dissect pro-survival mechanisms that cancer cells exploit during oncogenic transformation. HeLa cells, driven by HPV-18 oncogenes, rely on anti-apoptotic programs to evade cell death, and GRINA likely contributes to this resistance. Thus, disrupting GRINA in this background sensitizes cells to ER stress and apoptosis-inducing agents, enabling studies of drug resistance, calcium-dependent cell death, and neuroprotective pathways. This model is particularly valuable for examining the interplay between viral oncoproteins and cellular stress responses, shedding light on therapeutic vulnerabilities in cervical adenocarcinoma and beyond.
Typical research applications encompass apoptosis profiling (caspase-3/7 activity assays, Annexin V/PI flow cytometry), ER stress analysis (RT-qPCR for GRP78/BiP and CHOP), and calcium imaging with Fluo-4 AM to monitor store-operated calcium entry. Co-immunoprecipitation experiments can confirm physical interactions between GRINA and BAX or IP3 receptors, while MTT and clonogenic assays assess cell viability and proliferation. Additional uses include investigating neuroprotective signaling, ischemic injury mechanisms, and connections to neurodegenerative disorders such as Alzheimer??s disease. For further technical support or to inquire about custom modifications, please contact Ascent Research.
