A Processive Protein Chimera Introduces Mutations across Defined DNA Regions In Vivo
Name
nihms-987007.pdf
Description
Accepted version
Size
1.04 MB
Format
Adobe PDF
Checksum (MD5)
4a9e971670b1f60221773f4a3df8e238
Author(s)
Moore, Christopher Lawrence
Papa, Louis John
Shoulders, Matthew D.
Date Issued
July 2018
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Citation
Moore, Christopher L. et al. "A Processive Protein Chimera Introduces Mutations across Defined DNA Regions In Vivo." Journal of the American Chemical Society 140, 37 (July 2018): 11560–11564 © 2018 American Chemical Society
Version
Author's final manuscript
Abstract
Laboratory time scale evolution in vivo relies on the generation of large, mutationally diverse gene libraries to rapidly explore biomolecule sequence landscapes. Traditional global mutagenesis methods are problematic because they introduce many off-target mutations that are often lethal and can engender false positives. We report the development and application of the MutaT7 chimera, a potent and highly targeted in vivo mutagenesis agent. MutaT7 utilizes a DNA-damaging cytidine deaminase fused to a processive RNA polymerase to continuously direct mutations to specific, well-defined DNA regions of any relevant length. MutaT7 thus provides a mechanism for in vivo targeted mutagenesis across multi-kb DNA sequences. MutaT7 should prove useful in diverse organisms, opening the door to new types of in vivo evolution experiments.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/jacs.8b04001
