The ANG Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that targets the human ANG gene, encoding angiogenin. This loss-of-function model enables detailed study of angiogenin??s functions in tRNA cleavage, stress adaptation, and angiogenic signaling. The polyclonal nature provides a heterogeneous allele pool, offering a robust and reproducible tool that avoids clonal artifacts. It is suitable for a broad range of biochemical and cell-based assays.
The host cell line, HEK293T, is a widely used human embryonic kidney epithelial cell line that stably expresses the SV40 large T-antigen. Derived from HEK293 cells transformed with sheared adenovirus 5 DNA, HEK293T cells are known for their high transfection efficiency and capacity for recombinant protein production. Their epithelial nature and robust growth characteristics make them a versatile platform for studying intracellular signaling, gene regulation, and cellular stress mechanisms. The expression of SV40 large T-antigen supports episomal replication of plasmids containing the SV40 origin, facilitating gene delivery and protein expression studies.
Angiogenin (ANG) is a ribonuclease A family member with dual roles: extracellularly it promotes angiogenesis, while intracellularly it cleaves tRNA to generate tiRNAs under stress. Its activation is triggered by hypoxia (HIF-1??), cytokines (IL-1??, TNF-??), and growth factors (VEGF, bFGF). After internalization, ANG translocates to the nucleus and produces tiRNAs that repress translation and aid stress adaptation. Extracellular ANG binds cell surface receptors, activating the PI3K/Akt/mTOR cascade to stimulate endothelial cell proliferation and migration. ANG activity is inhibited by RNH1, and its localization is influenced by interactions with actin and TAF15. Downstream targets include rRNA transcription and cell migration pathways.
In the HEK293T background, the ANG knockout model allows dissection of angiogenin??s intracellular roles independently of endothelial contexts. These cells facilitate study of nuclear translocation, tiRNA biogenesis, and the impact of ANG loss on stress granule dynamics and translational repression. Co-culture or conditioned media experiments with endothelial cells can model paracrine angiogenic signaling. The knockout provides a clean background for reconstitution with ANG mutants to explore structure?Cfunction relationships relevant to cancer and neurodegenerative diseases.
This knockout model is ideal for studies on cancer cell migration and invasion, where ANG overexpression correlates with metastasis, and for investigating neuroprotection mechanisms in ALS, where ANG loss-of-function mutations are linked to disease. It also enables analysis of stress-induced tRNA fragment biology via RNA-seq and small RNA profiling. Typical assays include Western blotting for ANG, tube formation assays, tRNA cleavage assays, cell proliferation and migration assays, PI3K/Akt phospho-signaling analysis, and immunofluorescence for nuclear translocation. By eliminating endogenous angiogenin, this polyclonal knockout cell population provides a clean system for functional studies. For further details, contact Ascent Research.
