In vivo hypermutation and continuous evolution

Molina, Rosana S.; Rix, Gordon; Mengiste, Amanuella A.; Álvarez, Beatriz; Seo, Daeje; Chen, Haiqi; Hurtado, Juan E.; Zhang, Qiong; García-García, Jorge Donato; Heins, Zachary J.; Almhjell, Patrick J.; Arnold, Frances H.; Khalil, Ahmad S.; Hanson, Andrew D.; Dueber, John E.; Schaffer, David V.; Chen, Fei; Kim, Seokhee; Fernández, Luis Ángel; Shoulders, Matthew D.; Liu, Chang C.

doi:10.1038/s43586-022-00119-5

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In vivo hypermutation and continuous evolution

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Abstract

Directed evolution has revolutionized biomolecular engineering by applying cycles of mutation, amplification and selection to genes of interest (GOIs). However, classical directed evolution methods that rely on manually staged evolutionary cycles constrain the scale and depth of the evolutionary search that is possible. Here we describe genetic systems that achieve cycles of rapid mutation, amplification and selection fully inside living cells, enabling the continuous evolution of GOIs as cells grow. These systems advance the scale, evolutionary search depth, ease and overall power of directed evolution and access important new areas of protein evolution and engineering.

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59 BASIC BIOLOGICAL SCIENCES experimental evolution genetic engineering

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Molina, Rosana S., Rix, Gordon, Mengiste, Amanuella A., Álvarez, Beatriz, Seo, Daeje, Chen, Haiqi, Hurtado, Juan E., Zhang, Qiong, García-García, Jorge Donato, Heins, Zachary J., Almhjell, Patrick J., Arnold, Frances H., Khalil, Ahmad S., Hanson, Andrew D., Dueber, John E., Schaffer, David V., Chen, Fei, Kim, Seokhee, Fernández, Luis Ángel, Shoulders, Matthew D., Liu, Chang C.. 2022-05-19. In vivo hypermutation and continuous evolution. https://doi.org/10.1038/s43586-022-00119-5

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