Fluorescence-Activated Cell Sorting as a Tool for Recombinant Strain Screening
Book chapter, 2022

Metabolic engineering of microbial cells is the discipline of optimizing microbial metabolism to enable and improve the production of target molecules ranging from biofuels and chemical building blocks to high-value pharmaceuticals. The advances in genetic engineering have eased the construction of highly engineered microbial strains and the generation of genetic libraries. Intracellular metabolite-responsive biosensors facilitate high-throughput screening of these libraries by connecting the levels of a metabolite of interest to a fluorescence output. Fluorescent-activated cell sorting (FACS) enables the isolation of highly fluorescent single cells and thus genotypes that produce higher levels of the metabolite of interest. Here, we describe a high-throughput screening method for recombinant yeast strain screening based on intracellular biosensors and FACS.

Biosensors

Metabolic engineering

Strain engineering

Fluorescence

Library screening

FACS

Promoter library

Libraries

gRNA library

Microbiology

High-throughput screening

Author

Christos Skrekas

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Raphael Ferreira

Harvard Medical School

Florian David

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Methods in Molecular Biology

10643745 (ISSN) 1940-6029 (eISSN)

Vol. 2513 39-57

Subject Categories

Cell Biology

Medical Biotechnology

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1007/978-1-0716-2399-2_4

PubMed

35781199

More information

Latest update

7/12/2024