Understanding gene regulation is critical for developing novel therapeutic strategies. This study presents a new method for generating human cell lines that function as reporters of transcriptional activity using recombinant adeno-associated virus (rAAV)–mediated homologous recombination. To overcome rAAV cargo size limitations, an EGFP–luciferase fusion gene was employed as both a selectable marker and transcriptional reporter. EGFP-based selection leveraged endogenous gene activation conditions to identify correctly targeted alleles. Using this approach, primary human fibroblast lines were generated in which EGFP–luciferase expression is driven by the endogenous c-Myc oncogene, providing a powerful tool for studying transcriptional regulation in human cells.
