Microbial Transcriptomics

One-Sentence Definition

Microbial transcriptomics (RNA-seq) measures which genes a microbe is expressing under a given condition, revealing regulation that genome sequence alone cannot show.

Simple Explanation

The genome says what a cell can do; the transcriptome says what it is doing right now.

Detailed Scientific Explanation

Workflow: RNA extraction → rRNA depletion (bacteria lack polyA selection) → library → sequencing → QC → mapping/pseudo-alignment → counts → normalization (TPM/CPM) → differential expression (DESeq2, edgeR).

Specialized variants:

  • Dual RNA-seq — pathogen and host transcripts simultaneously
  • Single-cell / scRNA-seq of host immune cells during infection
  • Term-seq / dRNA-seq — transcript boundaries, operons
  • Nanopore direct RNA — native modifications

Interpretation: fold changes need biological replicates; expression ≠ protein level (Proteomics and MALDI Bioinformatics).

Mechanism

Counting reads per feature, modeling counts with negative binomial distributions, and controlling false discovery across thousands of genes.

Clinical Importance

  • Explains induced resistance (efflux pump upregulation, ampC induction) that gene detection misses
  • Host response signatures for sepsis discrimination feed AI Diagnostics in Microbiology

Research Importance

  • Regulatory networks, stress responses, biofilm and persister states

Diagnostic Relevance

  • Host transcriptomic signatures (bacterial vs viral) are an emerging diagnostic class

AMR Relevance

Active Recall Questions

  1. Why can’t you use polyA selection for bacterial mRNA?
  2. Which resistance mechanisms are visible only in expression data?
  3. What is dual RNA-seq?

Connections