The ANG Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, engineered to disrupt the human ANG gene encoding angiogenin. This heterogeneous polyclonal pool contains a variety of loss-of-function mutations introduced across the target locus, enabling functional studies of ANG without clonal isolation. The gene disruption is achieved through CRISPR/Cas9-mediated genome editing, resulting in abrogation of angiogenin protein expression and its associated cellular activities, providing a robust model for investigating angiogenin-dependent processes.
The HeLa host cell line is an immortalized human cervical adenocarcinoma cell line derived from an HPV18-positive tumor. As one of the most widely utilized human cell lines in biomedical research, HeLa cells offer a well-characterized epithelial cancer background with robust growth characteristics and genetic tractability. Their HPV-driven immortalization and dysregulated signaling pathways, including PI3K/Akt and NF-??B, make them particularly relevant for studying oncogenic mechanisms, angiogenesis, and stress responses, where angiogenin plays a key role.
Angiogenin (ANG) is a secreted ribonuclease that functions as a potent inducer of angiogenesis and cell proliferation. It is activated under hypoxic conditions via HIF-1?? and in response to inflammatory cytokines such as TNF-?? and IL-1??, and growth factors like VEGF and bFGF. ANG signals through a putative 170-kDa cell surface receptor, leading to activation of the PI3K/Akt/mTOR and ERK1/2 pathways, which promote rRNA transcription and cell survival. Under cellular stress, ANG translocates to the cytoplasm and cleaves tRNAs to produce tRNA-derived stress-induced small RNAs (tiRNAs), which facilitate stress granule assembly through interaction with G3BP1 and TIA-1, resulting in translational repression. ANG also interacts with actin and ribosome subunits, linking extracellular signaling to ribosome biogenesis and stress adaptation.
In the HeLa cervical cancer background, perturbation of ANG expression allows direct interrogation of its contributions to tumor cell proliferation, angiogenic signaling, and stress granule dynamics. Given the HPV18-driven activation of PI3K/Akt and NF-??B cascades, ANG knockout in this model dissects convergent signaling nodes that drive oncogenic phenotypes. This system is particularly valuable for studying the crosstalk between angiogenin-mediated tRNA cleavage and stress responses in cancer cells, as well as evaluating how loss of ANG impacts downstream targets like Akt and ERK1/2 within the context of an established cervical adenocarcinoma model.
Researchers can employ these ANG knockout HeLa polyclonal cells in a wide array of functional assays, including western blotting for angiogenin protein levels, RT-qPCR for mRNA quantification, RNase activity assays, cell proliferation and tube formation assays, immunofluorescence detection of stress granules, and tRNA fragment analysis via RNA-seq. These cells are ideal for studying angiogenin-mediated signaling in both cancer biology and neurodegenerative diseases such as amyotrophic lateral sclerosis and Parkinson’s disease, as well as for high-throughput screening of angiogenin inhibitors or activators. For further technical details and availability, please contact Ascent Research.
