A high-throughput and highly automated genotoxicity screening assay

Area of Science:

• Toxicology
• Genomics
• Biomarker Development

Background:

• Chemical safety assessments face challenges due to the increasing number of compounds requiring evaluation.
• In vitro screening using toxicogenomic biomarkers is an internationally endorsed first-tier approach for chemical assessment.
• A DNA damage-inducing (DDI) responsive transcriptomic biomarker, TGx-DDI, was previously developed for genotoxicity testing.

Purpose of the Study:

• To develop a highly automated, multiplexed, and high-throughput genotoxicity testing assay using Nanostring nCounter Plexset technology.
• To enhance the screening efficiency of the TGx-DDI biomarker for genotoxicity testing.
• To streamline chemical screening and assessment pipelines.

Main Methods:

• Development of the TGx-DDI Plexset assay utilizing Nanostring nCounter Plexset technology.
• Implementation of a highly automated and multiplexed workflow for genotoxicity testing.
• Demonstration of high-throughput capability by omitting concentration determination and RNA extraction steps.

Main Results:

• The TGx-DDI Plexset assay increased screening efficiency eight-fold compared to standard Nanostring nCounter technology.
• Hands-on time was significantly decreased with the automated, multiplexed assay.
• Assay performance, including specificity and sensitivity of TGx-DDI, was maintained without concentration determination and RNA extraction.

Conclusions:

• The TGx-DDI Plexset assay offers a simple, highly automated, multiplexed, and high-throughput pipeline for genotoxicity testing.
• This assay can be widely adopted for efficient chemical screening and safety assessment.
• The developed assay addresses the need for faster and more efficient regulatory evaluations of chemical compounds.