Strategies for in vitro engineering of the translation machinery.

Michael J Hammerling, Antje Krüger, Michael C Jewett

Journal: Nucleic acids research 2020;48(3):1068-1083

PMID: 31777928

Abstract

Engineering the process of molecular translation, or protein biosynthesis, has emerged as a major opportunity in synthetic and chemical biology to generate novel biological insights and enable new applications (e.g. designer protein therapeutics). Here, we review methods for engineering the process of translation in vitro. We discuss the advantages and drawbacks of the two major strategies-purified and extract-based systems-and how they may be used to manipulate and study translation. Techniques to engineer each component of the translation machinery are covered in turn, including transfer RNAs, translation factors, and the ribosome. Finally, future directions and enabling technological advances for the field are discussed.

© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

Address: Department of Chemical and Biological Engineering, Center for Synthetic Biology, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208, USA.
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