Single-Cell RNA Sequencing in Yeast Using the 10× Genomics Chromium Device

Area of Science:

• Microbiology
• Molecular Biology
• Genomics

Background:

• Single-cell RNA sequencing (scRNA-seq) is crucial for cellular physiology but challenging for microorganisms.
• Yeast presents unique difficulties for scRNA-seq due to cell wall rigidity, small size, and low RNA content.

Purpose of the Study:

• To adapt and optimize the 10× Genomics scRNA-seq protocol for the yeast Saccharomyces cerevisiae.
• To enable high-throughput transcriptomic analysis of individual yeast cells.

Main Methods:

• Incorporation of Zymolyase, a cell wall-degrading enzyme, into the droplet formation step.
• Utilizing the 10× Genomics platform with modifications for yeast cell lysis.
• Developing specific downstream data analysis pipelines and R-scripts for yeast scRNA-seq data.

Main Results:

• Successful adaptation of the scRNA-seq protocol for efficient lysis of Saccharomyces cerevisiae.
• Demonstrated compatibility of the modified protocol with standard droplet-based scRNA-seq workflows.
• Provided a comprehensive analysis pipeline for yeast scRNA-seq data.

Conclusions:

• The developed protocol effectively addresses the technical challenges of applying scRNA-seq to yeast.
• This adaptation expands the utility of scRNA-seq for microbial research, particularly in yeast.
• Facilitates detailed investigation of yeast cellular heterogeneity and gene expression patterns.