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A chimeric viral platform for directed evolution in mammalian cells
Open accessPreprint — published as 10.1038/s41467-025-59438-2 · Paper page
Abstract
Directed evolution (DE) is a process of mutation and artificial selection to breed biomolecules with new or improved activity 1,2 . DE platforms are primarily prokaryotic or yeast-based, and stable mutagenic mammalian systems have been challenging to establish and apply 3 . To this end, we developed PROTein Evolution Using Selection (PROTEUS), a new platform that uses chimeric virus-like vesicles (VLVs) to enable extended mammalian DE campaigns without loss of system integrity. This platform is stable and can generate sufficient diversity for DE in mammalian systems. Using PROTEUS, we altered the doxycycline responsiveness of tetracycline-controlled transactivators, generating a more sensitive TetON-4G tool for gene regulation. PROTEUS is also compatible with intracellular nanobody evolution, and we use it to design a DNA damage-responsive anti-p53 nanobody. Overall, PROTEUS is an efficient and stable platform to direct evolution of biomolecules within mammalian cells.
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Highly accurate protein structure prediction with AlphaFold
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Metadata
- Authors
- Cole, A. J., Denes, C. E., Moreno, C. L., Dobson, T., Hesselson, D., & Neely, G. G.
- Affiliations
- The University of Sydney (AU)
- Corresponding author
- Hesselson, Daniel
- Neely, G. Gregory
- ORCID
- Venue
- Nature Communications
- Publisher
- openRxiv
- Published
- 2024-04-20
- Volume
- 16
- Issue
- 1
- Pages
- 4250-4250
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC12059018
- OpenAlex
- W4410139679
- Cited by
- 7
- References
- 1
- Citations by year
- 2026: 5 · 2025: 2
- FWCI
- 4.09
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Nature Communications (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- NSW Ministry of Health
- Tour de Cure
- Medical Research Council
- National Health and Medical Research Council (1158164, 1158165, 1185002, 1107514)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Evolution and Genetic Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- directed evolution, mammalian cells, tetracycline-controlled transactivator, DNA damage response, gene expression regulation, protein evolution
- MeSH terms
- Animals, Humans, Tumor Suppressor Protein p53, Directed Molecular Evolution, HEK293 Cells, Single-Domain Antibodies
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