High Throughput Yeast Strain Phenotyping with Droplet-Based RNA Sequencing.
J Vis Exp
; (159)2020 05 21.
Article
em En
| MEDLINE
| ID: mdl-32510483
ABSTRACT
The powerful tools available to edit yeast genomes have made this microbe a valuable platform for engineering. While it is now possible to construct libraries of millions of genetically distinct strains, screening for a desired phenotype remains a significant obstacle. With existing screening techniques, there is a tradeoff between information output and throughput, with high-throughput screening typically being performed on one product of interest. Therefore, we present an approach to accelerate strain screening by adapting single cell RNA sequencing to isogenic picoliter colonies of genetically engineered yeast strains. To address the unique challenges of performing RNA sequencing on yeast cells, we culture isogenic yeast colonies within hydrogels and spheroplast prior to performing RNA sequencing. The RNA sequencing data can be used to infer yeast phenotypes and sort out engineered pathways. The scalability of our method addresses a critical obstruction in microbial engineering.
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Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
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Esferoplastos
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RNA Fúngico
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Engenharia Genética
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Análise de Sequência de RNA
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Ensaios de Triagem em Larga Escala
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article