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Lineage Tracing and Single-Cell RNA-seq in C. elegans to Analyze Transgenerational Epigenetic Phenotypes Inherited
Juan D Rodriguez1, David J Katz2
1Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 18, 2023
Summary
Transgenerational epigenetic inheritance in C. elegans can now be studied in embryos. This guide details automated lineage tracing and single-cell RNA-seq for analyzing germline effects on development.
Area of Science:
- Developmental Biology
- Epigenetics
- Genomics
Background:
- Transgenerational epigenetic inheritance, where traits pass across generations without DNA changes, is increasingly observed.
- Caenorhabditis elegans (C. elegans) is a key model for studying this phenomenon due to its short generation time and hermaphroditic reproduction.
- The impact of germline epigenetic inheritance on C. elegans embryogenesis remains poorly understood.
Purpose of the Study:
- To provide a practical guide for utilizing automated lineage tracing and single-cell RNA-sequencing (scRNA-seq) in C. elegans embryos.
- To enable the study of how transgenerational epigenetic inheritance from the germline influences embryogenesis.
- To overcome technical challenges hindering the adoption of these techniques in the research community.
Main Methods:
- Automated lineage tracing of cell development in C. elegans embryos.
- Single-cell RNA sequencing (scRNA-seq) to analyze gene expression at the cellular level.
- Integration of lineage tracing with scRNA-seq to map developmental trajectories and cell fates.
Main Results:
- Demonstration of a practical protocol for combining automated lineage tracing and scRNA-seq in C. elegans embryos.
- Establishment of methods to analyze the effects of germline transgenerational epigenetic inheritance on developmental timing and cell fate.
- Facilitation of high-resolution analysis of epigenetic inheritance during early development.
Conclusions:
- Automated lineage tracing coupled with scRNA-seq offers a powerful approach to investigate transgenerational epigenetic inheritance in C. elegans embryogenesis.
- This methodology can elucidate the mechanisms by which epigenetic information is transmitted and impacts development across generations.
- The provided guide aims to increase the accessibility and application of these advanced techniques for studying developmental epigenetics.
