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Nanopore based analyses of genome-wide DNA methylation profiles through Tribolium castaneum development

Epigenetic processes are critical during development and are required to maintain a balanced state of gene expression in a tissue and cell specific manner. Among epigenetic modifications DNA methylation is one of the most widely studied and often in combination with other chromatin modifications can provide valuable insight into processes impacting development. Recent advances in Nanopore sequencing technology show that detection of different DNA modifications is possible through the analysis of raw nanopore signals. This novel sequencing technology circumvents problems of traditional methods and allows detection of long-read whole-genome methylation patterns. The red flour beetle Tribolium castaneum represents an important model system for insect functional genetics. It is used for studying insect typical development, the evolution of development and for research on metabolism and pest control. Here, we profiled the genome-wide DNA methylation status of T. castaneum in four developmental stages ; eggs, larvae, male and female pupae and adults. First, we generated 10-18 Gb of raw sequence data per sample to provide sufficient depth of over 30X. Both 6mA and 5mC modifications were detected on raw sequence data using the neural network-based software Megalodon. Preliminary results show the highest methylation dynamics of both 5mC and 6mA in genes related to the formation of the cuticle, odorant-binding proteins and olfactory receptors.

Tribolium ; Nanopore ; Sequencing ; DNA methylation

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