Duplex Sequencing - Future of mutagenicity assessment

With so much discussion around Limits and AI calculations, I want to debate for a moment to share information about ‘Duplex Sequencing’. After the recent safety discussion at the Hesi/FDA meeting, I went down a rabbit hole to understand some of the proposed assays and testing that can be used to evaluate the safety or mutagenicity potential in Nitrosamines Impurities.

Duplex Sequencing - I would like to share couple peer-reviewd articles that provide a perspective about duplex sequencing and how this can be used to assess mutaginicity.


Does anyone has experience performing duplex sequencing (DS) methodology in wild-type rodent tissues, without the need for transgenic animals? Are there any reliable CRO’s for performing DS analyses?

I understand one of the beneficts of DS is that you do not need transgenic animals + the cost compared to a TGRA. If I am not mistaken DS is a propietary methodology, at least part of the materials.

TwinStrand Duplex Sequencing Technology (twinstrandbio.com)

The only caveat, is that there is no OECD guideline for it and that may take years. Therefore, to avoid heterogenicity, a common “procedure” done by sponsors should be available first.


Thank you for your answer. I agree.

I believe DS is just one of the next-generation sequencing method and TwinStrand Biosciences actually developed a DS-based mutagenesis assay. However, except for the TwinStrand I could not come across any other CRO performing duplex sequencing for which I believe is a promising tool for cost-effective, accurate and time efficient method.