A chimeric viral platform for directed evolution in mammalian cells
Abstract Directed evolution is a process of mutation and artificial selection to breed biomolecules with new or improved activity. Directed evolution platforms are primarily prokaryotic or yeast-based, and stable mammalian systems have been challenging to establish and apply. To this end, we develop...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-05-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-59438-2 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850278115279896576 |
|---|---|
| author | Alexander J. Cole Christopher E. Denes Cesar L. Moreno Lise Hunault Thomas Dobson Daniel Hesselson G. Gregory Neely |
| author_facet | Alexander J. Cole Christopher E. Denes Cesar L. Moreno Lise Hunault Thomas Dobson Daniel Hesselson G. Gregory Neely |
| author_sort | Alexander J. Cole |
| collection | DOAJ |
| description | Abstract Directed evolution is a process of mutation and artificial selection to breed biomolecules with new or improved activity. Directed evolution platforms are primarily prokaryotic or yeast-based, and stable mammalian systems have been challenging to establish and apply. To this end, we develop PROTein Evolution Using Selection (PROTEUS), a platform that uses chimeric virus-like vesicles to enable extended mammalian directed evolution campaigns without loss of system integrity. This platform is stable and can generate sufficient diversity for directed evolution in mammalian systems. Using PROTEUS, we alter the doxycycline responsiveness of tetracycline-controlled transactivators, generating a more sensitive TetON-4G tool for gene regulation with mammalian-specific adaptations. PROTEUS is also compatible with intracellular nanobody evolution, and we use it to evolve a DNA damage-responsive anti-p53 nanobody. Overall, PROTEUS is an efficient and stable platform to direct evolution of biomolecules within mammalian cells. |
| format | Article |
| id | doaj-art-91ae947ea7a04f6ab29f4f96a6b2781e |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-91ae947ea7a04f6ab29f4f96a6b2781e2025-08-20T01:49:37ZengNature PortfolioNature Communications2041-17232025-05-0116111310.1038/s41467-025-59438-2A chimeric viral platform for directed evolution in mammalian cellsAlexander J. Cole0Christopher E. Denes1Cesar L. Moreno2Lise Hunault3Thomas Dobson4Daniel Hesselson5G. Gregory Neely6Centenary Institute and Faculty of Medicine and Health, Charles Perkins Centre, The University of SydneyThe Dr. John and Anne Chong Lab for Functional Genomics, School of Life & Environmental Sciences, Charles Perkins Centre, The University of SydneyThe Dr. John and Anne Chong Lab for Functional Genomics, School of Life & Environmental Sciences, Charles Perkins Centre, The University of SydneyCentenary Institute and Faculty of Medicine and Health, Charles Perkins Centre, The University of SydneyCentenary Institute and Faculty of Medicine and Health, Charles Perkins Centre, The University of SydneyCentenary Institute and Faculty of Medicine and Health, Charles Perkins Centre, The University of SydneyThe Dr. John and Anne Chong Lab for Functional Genomics, School of Life & Environmental Sciences, Charles Perkins Centre, The University of SydneyAbstract Directed evolution is a process of mutation and artificial selection to breed biomolecules with new or improved activity. Directed evolution platforms are primarily prokaryotic or yeast-based, and stable mammalian systems have been challenging to establish and apply. To this end, we develop PROTein Evolution Using Selection (PROTEUS), a platform that uses chimeric virus-like vesicles to enable extended mammalian directed evolution campaigns without loss of system integrity. This platform is stable and can generate sufficient diversity for directed evolution in mammalian systems. Using PROTEUS, we alter the doxycycline responsiveness of tetracycline-controlled transactivators, generating a more sensitive TetON-4G tool for gene regulation with mammalian-specific adaptations. PROTEUS is also compatible with intracellular nanobody evolution, and we use it to evolve a DNA damage-responsive anti-p53 nanobody. Overall, PROTEUS is an efficient and stable platform to direct evolution of biomolecules within mammalian cells.https://doi.org/10.1038/s41467-025-59438-2 |
| spellingShingle | Alexander J. Cole Christopher E. Denes Cesar L. Moreno Lise Hunault Thomas Dobson Daniel Hesselson G. Gregory Neely A chimeric viral platform for directed evolution in mammalian cells Nature Communications |
| title | A chimeric viral platform for directed evolution in mammalian cells |
| title_full | A chimeric viral platform for directed evolution in mammalian cells |
| title_fullStr | A chimeric viral platform for directed evolution in mammalian cells |
| title_full_unstemmed | A chimeric viral platform for directed evolution in mammalian cells |
| title_short | A chimeric viral platform for directed evolution in mammalian cells |
| title_sort | chimeric viral platform for directed evolution in mammalian cells |
| url | https://doi.org/10.1038/s41467-025-59438-2 |
| work_keys_str_mv | AT alexanderjcole achimericviralplatformfordirectedevolutioninmammaliancells AT christopheredenes achimericviralplatformfordirectedevolutioninmammaliancells AT cesarlmoreno achimericviralplatformfordirectedevolutioninmammaliancells AT lisehunault achimericviralplatformfordirectedevolutioninmammaliancells AT thomasdobson achimericviralplatformfordirectedevolutioninmammaliancells AT danielhesselson achimericviralplatformfordirectedevolutioninmammaliancells AT ggregoryneely achimericviralplatformfordirectedevolutioninmammaliancells AT alexanderjcole chimericviralplatformfordirectedevolutioninmammaliancells AT christopheredenes chimericviralplatformfordirectedevolutioninmammaliancells AT cesarlmoreno chimericviralplatformfordirectedevolutioninmammaliancells AT lisehunault chimericviralplatformfordirectedevolutioninmammaliancells AT thomasdobson chimericviralplatformfordirectedevolutioninmammaliancells AT danielhesselson chimericviralplatformfordirectedevolutioninmammaliancells AT ggregoryneely chimericviralplatformfordirectedevolutioninmammaliancells |