The IFI27 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the IFI27 gene in the HeLa human cervical adenocarcinoma cell line. This loss-of-function model eliminates IFI27 protein expression, enabling detailed investigation of its role in apoptosis and interferon signaling. The polyclonal format provides a heterogeneous knockout pool suitable for studies where genetic diversity is advantageous, without the selection bias of single-cell clones.
HeLa cells are an HPV18-positive, immortalized epithelial line originating from cervical adenocarcinoma. They exhibit robust growth, well-defined interferon signaling pathways, and are extensively used in cancer research. The cervical cancer origin makes them particularly relevant for studying tumor cell apoptosis and virus-host interactions.
IFI27 is an interferon-stimulated gene that encodes a pro-apoptotic protein. Its transcription is driven by type I interferons (IFN-??/??) through the JAK-STAT cascade: binding to IFNAR1/2 receptors activates JAK1 and TYK2, which phosphorylate STAT1 and STAT2, leading to ISGF3 complex formation with IRF9 and subsequent binding to ISRE promoter elements. IFI27 protein subsequently interacts with BCL2 family members, including BAX, BCL2, and BCL-XL, promoting mitochondrial outer membrane permeabilization, cytochrome c release, and activation of caspase-9 and caspase-3 to execute apoptosis.
In the HeLa background, loss of IFI27 provides insights into how interferon-induced apoptosis is regulated in cancer cells, where HPV18 oncoproteins E6 and E7 already disrupt p53 and Rb tumor suppressor pathways. This model is valuable for studying resistance to interferon-based therapies and immune-mediated cytotoxicity, as well as the interplay between viral oncogenesis and mitochondrial cell death.
Applications include RT-qPCR and Western blotting for IFI27 expression analysis, apoptosis assays (annexin V staining, caspase-3/9 activation, mitochondrial membrane potential), co-immunoprecipitation for protein interactions, and immunofluorescence localization. Viral infection models benefit from this knockout to delineate IFI27-dependent defense mechanisms. For further information, please contact Ascent Research.
