The HERC5 Knouckout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HERC5 gene in the A-549 lung adenocarcinoma line. HERC5 encodes an interferon-induced E3 ubiquitin ligase that mediates ISG15 conjugation. Disruption of HERC5 eliminates ISGylation activity, providing a loss-of-function model for studying innate immunity. The polyclonal format avoids clonal artifacts and maintains heterogeneous cellular responses.
The A-549 cell line originates from a lung adenocarcinoma of a 58-year-old male and serves as a canonical model for non-small cell lung cancer (NSCLC). These epithelial cells of alveolar type II origin are widely used in cancer research and antiviral studies due to their intact interferon signaling pathways.
HERC5 functions as an ISG15 E3 ligase, catalyzing ISGylation of target proteins. Its expression is induced by type I interferons (IFN-??/??) via transcription factors IRF3, IRF7, and STAT1. Interacting partners include UBE2L6 and UBA7, forming the ISGylation conjugation machinery. HERC5 targets viral proteins, IRF3, STAT1, and cytosolic DNA sensors, thereby modulating key innate immune pathways such as RIG-I/MDA5/cGAS-STING-TBK1/IKK??-IRF3 and IFNAR-JAK-STAT signaling. Through ISGylation, HERC5 regulates protein function and antiviral defense.
In A-549 cells, HERC5 knockout abrogates ISGylation, enabling precise dissection of this modification in lung cancer biology. This model is relevant for studying oncolytic virus therapy, as ISGylation can restrict viral replication. It also facilitates investigation of host-pathogen interactions for viruses like influenza and hepatitis, and explores links to autoimmune disorders driven by interferon dysregulation.
Typical applications include western blotting for ISG15 conjugates and free ISG15, RT-qPCR of ISGs, antiviral resistance assays, co-immunoprecipitation for ISGylated proteins, immunofluorescence of viral replication, and flow cytometry for MHC-I expression. These cells are suited for innate immunity, cancer immunology, and oncolytic virotherapy research. Contact Ascent Research for further information.
