Bacterial Single Cell Whole Transcriptome Amplification in Microfluidic Platform Shows Putative Gene Expression...

Area of Science:

• Microbiology
• Genomics
• Molecular Biology

Background:

• Single cell RNA sequencing (scRNA-seq) enables analysis of individual cell transcriptomes, but its application to microbial cells is challenging due to low RNA input.
• Existing amplification methods for microbial scRNA-seq often introduce contamination and bias.
• Porphyromonas somerae is a microbe associated with endometrial cancer, making its gene expression patterns of interest.

Purpose of the Study:

• To develop and validate a method for single-cell whole transcriptome amplification and sequencing in microbial cells.
• To investigate gene expression heterogeneity within a population of Porphyromonas somerae.
• To demonstrate the utility of scRNA-seq for studying microbial gene regulation.

Main Methods:

• Coupling a microfluidic platform with multiple displacement amplification (MDA) technology.
• Performing whole transcriptome amplification on single microbial cells.
• Sequencing the amplified RNA from single Porphyromonas somerae cells.

Main Results:

• Successfully achieved single-cell whole transcriptome amplification and sequencing for Porphyromonas somerae.
• Demonstrated distinct gene expression regulation across individual microbial cells.
• Provided evidence for widespread gene expression heterogeneity in bacterial populations.

Conclusions:

• The developed microfluidic-MDA approach is effective for microbial scRNA-seq.
• Single microbial cells exhibit significant variations in gene expression.
• This technique can unveil crucial insights into microbial biology and disease associations.