@inbook{5bb0900c7dd64b3fbbdc9c141f0cecb1,
title = "Emulsion-based directed evolution of enzymes and proteins in yeast",
abstract = "Emulsion-based selections are a unique type of directed evolution method that overcome common bottlenecks associated with purely in vivo selections. For example, emulsions including cell-free translation machinery can be useful for expression of toxic genes. However, not all cell types can efficiently produce protein in vitro, for example, the eukaryotic microbe Saccharomyces cerevisiae. compartmentalized self replication (CSR) and compartmentalized partnered replication (CPR) are two emulsion-based selection schemes that leverage the advantages of both in vivo and in vitro selections by compartmentalizing cells in water-in-oil droplets. Previous implementations of these methods utilized bacterial hosts, which has limited the technology to the directed evolution of proteins that can be heterologously expressed in prokaryotic systems. To expand the repertoire of targets that can be evolved, we have adapted emulsion-based PCR selections to be compatible with a eukaryotic host.",
keywords = "Cellular compartmentalization, Directed evolution, Emulsion PCR, Emulsion droplets, Emulsions, Protein engineering, Screening, Selection",
author = "Gardner, {Elizabeth C.} and Watkins, {Ella J.} and Jimmy Gollihar and Ellington, {Andrew D.}",
note = "Funding Information: This work was supported by NSF SemiSynBio Award 1807369. Special thanks to Dr. Katy Kao for transcribing the first draft of the protocol. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
doi = "10.1016/bs.mie.2020.04.053",
language = "English (US)",
isbn = "9780128211496",
volume = "643",
series = "Methods in enzymology",
publisher = "Academic Press",
pages = "87--110",
editor = "Tawfik, {Dan S.}",
booktitle = "Enzyme Engineering and Evolution",
address = "United States",
}