Abstract
Stable mammalian cell lines with sustained transgene expression are indispensable for functional genomics and long-term genetic studies. However, transient approaches often suffer from inconsistent efficiency and expression instability across diverse cell types. This MiniResource addresses these challenges by providing a comprehensive synthesis of gene delivery strategies and optimized, systematic workflows for establishing robust cell lines via chemical transfection and viral transduction. By distilling complex procedures into accessible, standardized guidelines, this work serves as an essential practical resource for achieving uniform and durable expression in both commonly used immortalized cell lines and challenging primary or immune cell populations.