Single-cell RNA sequencing algorithms underestimate changes in transcriptional noise compared to single-molecule RNA imaging
2024

Understanding Transcriptional Noise in Cells

Sample size: 811 publication 10 minutes Evidence: moderate

Author Information

Author(s): Khetan Neha, Zuckerman Binyamin, Calia Giuliana P., Chen Xinyue, Garcia Arceo Ximena, Weinberger Leor S.

Primary Institution: Gladstone|UCSF Center for Cell Circuitry, University of California, San Francisco

Hypothesis

How can we best quantify genome-wide transcriptional noise?

Conclusion

The study finds that most single-cell RNA sequencing algorithms underestimate transcriptional noise changes compared to single-molecule RNA imaging.

Supporting Evidence

  • scRNA-seq validates that IdU orthogonally amplifies transcriptome-wide noise.
  • smFISH validates IdU-induced noise amplification for most genes.
  • scRNA-seq algorithms underestimate the fold change in noise compared to smFISH.
  • The underestimation of changes in noise holds even after extrinsic factor corrections.

Takeaway

This study looks at how noise in gene expression can change without affecting the average amount of gene product, and it shows that some methods for measuring this noise don't work as well as others.

Methodology

The study used single-cell RNA sequencing and single-molecule RNA fluorescence in situ hybridization to analyze transcriptional noise in human and mouse datasets.

Potential Biases

Potential biases from technical noise in scRNA-seq methods could affect the quantification of transcriptional noise.

Limitations

The study is limited by a single replicate in mouse embryonic stem cells and smaller sequencing depth in Jurkat cells.

Participant Demographics

The study involved mouse embryonic stem cells and human Jurkat T lymphocytes.

Statistical Information

P-Value

p<10−17

Statistical Significance

p<10−17

Digital Object Identifier (DOI)

10.1016/j.crmeth.2024.100933

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