Mammalian cell-derived protein therapeutic production has changed the landscape of human healthcare in the past two decades. The importance of protein therapeutics has motivated the search for more cost-effective and efficient cell lines capable of producing high quality protein products. The factors contributing to optimal producer cell lines are often complex, and not simply conferred by one gene or gene product, which makes an understanding of system-wide properties for better engineering of optimized cell lines essential. Genome-scale technologies (genomics, transcriptomics and proteomics) enable such engineering studies. However, the use of these technologies in cell culture engineering is still in its infancy. Here, we summarize current knowledge of cell properties important for the design of efficient protein-producing mammalian cell lines, and highlight relevant studies to-date that use genome-scale technologies in these cell systems. We also provide a focused review of relevant alternative and emerging technologies, which have seen limited use in cell culture engineering, but hold great potential for significant advancements in protein therapeutic production.