High throughput mutagenesis and screening for yeast engineering

• Type of Material: Article
• Authors: Kendreze Holland, Emory University; John Blazeck, Georgia Institute of Technology
• Language: English
• Date: 2022-12-27
• Publisher: BioMed Central
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2022
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1186/s13036-022-00315-7
• Title of Journal or Parent Work: Journal of Biological Engineering
• Volume: 16
• Grant/Funding Information: Kendreze Holland is supported by the National Science Foundation’s Graduate Research Fellowship Program. John Blazeck is supported by the Beckman Young Investigator Award from the Arnold and Mabel Beckman Foundation.
• Keywords: Cellular engineering; High throughput; Selection schemes; Genome-wide; Yeast-display; Directed evolution; S. cerevisiae; In vivo mutagenesis; Protein engineering; CRISPR-Cas9
• Research Categories: Engineering, Chemical; Engineering, Biomedical

Abstract

The eukaryotic yeast Saccharomyces cerevisiae is a model host utilized for whole cell biocatalytic conversions, protein evolution, and scientific inquiries into the pathogenesis of human disease. Over the past decade, the scale and pace of such studies has drastically increased alongside the advent of novel tools for both genome-wide studies and targeted genetic mutagenesis. In this review, we will detail past and present (e.g., CRISPR/Cas) genome-scale screening platforms, typically employed in the context of growth-based selections for improved whole cell phenotype or for mechanistic interrogations. We will further highlight recent advances that enable the rapid and often continuous evolution of biomolecules with improved function. Additionally, we will detail the corresponding advances in high throughput selection and screening strategies that are essential for assessing or isolating cellular and protein improvements. Finally, we will describe how future developments can continue to advance yeast high throughput engineering.