A key limitation of all directed evolution workflows performed in test tubes, bacteria, or yeast is that these approaches often yield products that fail to function when introduced into mammalian cells. Instead, functions evolved in simpler systems are often derailed in complex mammalian cells by off-target interactions, poor stability, inappropriate modification/localization, or many other serious problems. This frontier challenge could theoretically be addressed by leveraging the human cell itself as the design, engineering, and quality control factory for biomolecule discovery and optimization. In 2018, we reported the first rapid and efficient system for continuous directed evolution directly in living mammalian cells. In our approach, essentially a mammalian phage-assisted continuous evolution (mPACE) platform, the ability of a highly error-prone adenovirus to propagate is coupled to the activity of a biomolecule of interest within the human cell itself. The system is designed for safety, speed, and ease-of-use. Critically, mutagenesis, selection, and amplification all occur concurrently in this platform, maximizing the benefits of natural selection and the throughput of experiments. Here, we will discuss progress in the application of mammalian cell-based continuous directed evolution to the engineering of desired biomolecule functions in human cells. We believe this type of system, as it and those like it continue to improve, holds potential for major impact on the fields of biomolecule engineering and biomedical research, as well as important applications in evolutionary biology.

Continuous Directed Evolution in Mammalian Cells

ACS Fall 2020 Virtual Meeting

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