CaltechAUTHORS
Published May 19, 2022 | Version public
Journal Article Metadata-only

In vivo hypermutation and continuous evolution

Creators

  • 1. ROR icon University of California, Irvine
  • 2. ROR icon Massachusetts Institute of Technology
  • 3. ROR icon National Center for Biotechnology
  • 4. ROR icon Seoul National University
  • 5. ROR icon The University of Texas Southwestern Medical Center
  • 6. ROR icon University of California, Berkeley
  • 7. ROR icon Monterrey Institute of Technology and Higher Education
  • 8. ROR icon Boston University
  • 9. ROR icon California Institute of Technology
  • 10. ROR icon Harvard University
  • 11. ROR icon University of Florida
  • 12. ROR icon Innovative Genomics Institute
  • 13. ROR icon Lawrence Berkeley National Laboratory
  • 14. ROR icon Broad Institute

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.

Additional Information

The authors thank members of their groups for insightful discussions. This work was funded by National Institutes of Health (NIH) National Institute of General Medical Sciences (NIGMS) 1R35GM139513 (C.C.L.); NIH NIGMS 1R35GM136354 (M.D.S.); MIT Robert J Silbey Fellowship (A.A.M.); MIT School of Science Fund for Future of Science (A.A.M.); US Department of Energy, Office of Science, Basic Energy Sciences under Award DE-SC0020153 (A.D.H.); Innovative Genomics Institute and Laboratory for Genomics Research (J.E.H., J.E.D. and D.V.S.); UC Berkeley Miller Basic Research Fellowship (Q.Z.); NIH National Institute of Biomedical Imaging and Bioengineering (NIBIB) 1R01EB027793 (A.S.K.); Department of Defense (DoD) Vannevar Bush Faculty Fellowship N00014-20-1-2825 (A.S.K.); NIH NIGMS 1R01GM125887 (F.H.A.); and Ministerio de Ciencia e Innovación - Consejo Superior de Investigaciones Científicas (MICIN-CSIC) PTI + REC-EU SGL2103051 and EU Horizon 2020 research and innovation programme FET Open 965018-BIOCELLPHE (L.A.F.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH or other funding agencies.

Additional details

Identifiers

Eprint ID
118312
Resolver ID
CaltechAUTHORS:20221212-796368000.22

Funding

NIH
1R35GM139513
NIH
1R35GM136354
Massachusetts Institute of Technology (MIT)
Department of Energy (DOE)
DE-SC0020153
Miller Institute for Basic Research in Science
NIH
1R01EB027793
Vannever Bush Faculty Fellowship
N00014-20-1-2825
NIH
1R01GM125887
Ministerio de Ciencia e Innovación (MICINN)
SGL2103051
Consejo Superior de Investigaciones Científicas (CSIC)
European Research Council (ERC)
965018

Dates

Created
2023-01-14
Created from EPrint's datestamp field
Updated
2023-01-17
Created from EPrint's last_modified field

Caltech Custom Metadata

Caltech groups
Division of Biology and Biological Engineering (BBE)