Modern Approaches in Genetic Toxicology Research: Insights from Next-Generation-Based RNA Sequencing

Area of Science:

• Toxicogenomics
• Molecular Biology
• Bioinformatics

Background:

• New Approach Methods (NAMs) are crucial for assessing DNA damage.
• Transcriptomics offers high-throughput, high-content, and human-relevant analyses.
• Gene expression profiling reveals adverse and adaptive biological responses to chemical exposure.

Purpose of the Study:

• To detail the methodology for RNA sequencing and data analysis for genotoxicity studies.
• To identify biomarkers and signatures for genotoxicity assessment.
• To provide insights into Wet Lab and Dry Lab procedures for RNA-Seq.

Main Methods:

• RNA isolation, cDNA synthesis, adapter ligation, and library preparation.
• Next-Generation Sequencing (NGS) techniques, including single-read and paired-end sequencing.
• Bioinformatic analysis of RNA sequencing data using R-programming.

Main Results:

• RNA sequencing enables comprehensive analysis of gene expression, including mRNA, small RNAs, and long non-coding RNAs.
• The method allows for the detection of alternative splicing, gene fusions, and post-transcriptional modifications.
• Identification of potential biomarkers for genotoxicity is facilitated by transcriptomic profiling.

Conclusions:

• Transcriptomics-based RNA sequencing is a powerful tool for genotoxicity testing in human-derived cell systems.
• This approach provides a detailed understanding of cellular responses to chemical exposures.
• The described methodology supports the development of NAMs for toxicological assessments.