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Evolution of new nonantibody proteins via iterative somatic hypermutation
Open accessAbstract
B lymphocytes use somatic hypermutation (SHM) to optimize immunoglobulins. Although SHM can rescue single point mutations deliberately introduced into nonimmunoglobulin genes, such experiments do not show whether SHM can efficiently evolve challenging novel phenotypes requiring multiple unforeseeable mutations in nonantibody proteins. We have now iterated SHM over 23 rounds of fluorescence-activated cell sorting to create monomeric red fluorescent proteins with increased photostability and far-red emissions (e.g., 649 nm), surpassing the best efforts of structure-based design. SHM offers a strategy to evolve nonantibody proteins with desirable properties for which a high-throughput selection or viable single-cell screen can be devised.
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Cites
Imaging tri-fusion multimodality reporter gene expression in living subjects
Ray, P., De, A., Min, J.-J., Tsien, R. Y., & Gambhir, S. S. (2004) · Cancer Research · doi:10.1158/0008-5472.can-03-1816 · Cited by 329
Self-citation
Cited by
Lichtman, J. W., & Conchello, J.-A. (2005) · Nature Methods · doi:10.1038/nmeth817 · Cited by 1,742
Fluorescent Proteins and Their Applications in Imaging Living Cells and Tissues
Chudakov, D. M., Matz, M. V., Lukyanov, S., & Lukyanov, K. A. (2010) · Physiological Reviews · doi:10.1152/physrev.00038.2009 · Cited by 1,375
Advances in fluorescent protein technology
Shaner, N. C., Patterson, G. H., & Davidson, M. W. (2007) · Journal of Cell Science · doi:10.1242/jcs.005801 · Cited by 787
FLUORESCENCE MOLECULAR IMAGING
Ntziachristos, V. (2006) · Annual Review of Biomedical Engineering · doi:10.1146/annurev.bioeng.8.061505.095831 · Cited by 731
Directed Evolution: Methodologies and Applications
Wang, Y., Xue, P., Cao, M., Yu, T., Lane, S. T., & Zhao, H. (2021) · Chemical Reviews · doi:10.1021/acs.chemrev.1c00260 · Cited by 719
Metadata
- Authors
- Wang, L., Jackson, W. C., Steinbach, P. A., & Tsien, R. Y.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2004-11-19
- Volume
- 101
- Issue
- 48
- Pages
- 16745-16749
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2058211348
- Cited by
- 416
- References
- 1
- FWCI
- 13.13
- Type
- journal article
- Language
- en
- Open access
- green
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- Wang, L., Jackson, W. C., Steinbach, P. A., & Tsien, R. Y. (2004). Evolution of new nonantibody proteins via iterative somatic hypermutation. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0407752101
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