In vivo hypermutation and continuous evolution

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

doi:10.1038/s43586-022-00119-5

In vivo hypermutation and continuous evolution

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

Research output: Contribution to journal › Review article › peer-review

49 Scopus citations

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. 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.

Original language	English (US)
Article number	36
Journal	Nature Reviews Methods Primers
Volume	2
Issue number	1
DOIs	https://doi.org/10.1038/s43586-022-00119-5
State	Published - Dec 2022

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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Molina, R. S., Rix, G., Mengiste, A. A., Álvarez, B., Seo, D., Chen, H., Hurtado, J. E., Zhang, Q., García-García, J. D., Heins, Z. J., Almhjell, P. J., Arnold, F. H., Khalil, A. S., Hanson, A. D., Dueber, J. E., Schaffer, D. V., Chen, F., Kim, S., Fernández, L. Á., ... Liu, C. C. (2022). In vivo hypermutation and continuous evolution. Nature Reviews Methods Primers, 2(1), Article 36. https://doi.org/10.1038/s43586-022-00119-5

Molina, RS, Rix, G, Mengiste, AA, Álvarez, B, Seo, D, Chen, H, Hurtado, JE, Zhang, Q, García-García, JD, Heins, ZJ, Almhjell, PJ, Arnold, FH, Khalil, AS, Hanson, AD, Dueber, JE, Schaffer, DV, Chen, F, Kim, S, Fernández, LÁ, Shoulders, MD & Liu, CC 2022, 'In vivo hypermutation and continuous evolution', Nature Reviews Methods Primers, vol. 2, no. 1, 36. https://doi.org/10.1038/s43586-022-00119-5

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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. 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.",

author = "Molina, {Rosana S.} and Gordon Rix and Mengiste, {Amanuella A.} and Beatriz {\'A}lvarez and Daeje Seo and Haiqi Chen and Hurtado, {Juan E.} and Qiong Zhang and Garc{\'i}a-Garc{\'i}a, {Jorge Donato} and Heins, {Zachary J.} and Almhjell, {Patrick J.} and Arnold, {Frances H.} and Khalil, {Ahmad S.} and Hanson, {Andrew D.} and Dueber, {John E.} and Schaffer, {David V.} and Fei Chen and Seokhee Kim and Fern{\'a}ndez, {Luis {\'A}ngel} and Shoulders, {Matthew D.} and Liu, {Chang C.}",

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doi = "10.1038/s43586-022-00119-5",

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AU - Molina, Rosana S.

AU - Rix, Gordon

AU - Mengiste, Amanuella A.

AU - Álvarez, Beatriz

AU - Seo, Daeje

AU - Chen, Haiqi

AU - Hurtado, Juan E.

AU - Zhang, Qiong

AU - García-García, Jorge Donato

AU - Heins, Zachary J.

AU - Almhjell, Patrick J.

AU - Arnold, Frances H.

AU - Khalil, Ahmad S.

AU - Hanson, Andrew D.

AU - Dueber, John E.

AU - Schaffer, David V.

AU - Chen, Fei

AU - Kim, Seokhee

AU - Fernández, Luis Ángel

AU - Shoulders, Matthew D.

AU - Liu, Chang C.

PY - 2022/12

Y1 - 2022/12

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AB - 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. 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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In vivo hypermutation and continuous evolution

Abstract

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