Cell Line A549

A549 cells are a human alveolar epithelial cell line derived from lung adenocarcinoma and are widely used in virology, cancer biology, innate immunity, drug discovery, ultrastructural studies, and general molecular and cell biology. Their broad utility stems from their human origin, epithelial characteristics, and ease of genetic manipulation. To investigate virus-induced organelle remodeling and host-pathogen interactions at the subcellular level, we developed a panel of A549 cell lines in which 16 distinct organelles were endogenously tagged with GFP using CRISPR-Cas9 genome editing. Following fluorescence-activated cell sorting, all lines were assessed for post-revival viability and confirmed to be mycoplasma-free. To ensure imaging reproducibility across experimental workflows, the cells were validated under live-cell, methanol-fixed, and paraformaldehyde-fixed conditions.