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Detection of genetic variation and base modifications at base-pair resolution on both DNA and RNA

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Wang, Zhen | Maluenda, Jérôme | Giraut, Laurène | Vieille, Thibault | Lefevre, Andréas | Salthouse, David | Radou, Gaël | Moulinas, Rémi | Astete, Sandra | d'Avezac, Pol | Smith, Geoff | André, Charles | Allemand, Jean-François | Bensimon, David | Croquette, Vincent | Ouellet, Jimmy | Hamilton, Gordon

Edité par CCSD ; Nature Publishing Group -

International audience. Accurate decoding of nucleic acid variation is critical to understand the complexity and regulation of genome function. Here we use a single-molecule magnetic tweezer (MT) platform to identify sequence variation and map a range of important epigenetic base modifications with high sensitivity, specificity, and precision in the same single molecules of DNA or RNA. We have also developed a highly specific amplification-free CRISPR-Cas enrichment strategy to isolate genomic regions from native DNA. We demonstrate enrichment of DNA from both E. coli and the FMR1 5'UTR coming from cells derived from a Fragile X carrier. From these kilobase-length enriched molecules we could characterize the differential levels of adenine and cytosine base modifications on E. coli, and the repeat expansion length and methylation status of FMR1. Together these results demonstrate that our platform can detect a variety of genetic, epigenetic, and base modification changes concomitantly within the same single molecules.

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