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L'organisation 3D des chromosomes synthétiques de levure

Abstract : The international project Sc2.0 started 10 years ago by the Pr. Jef Boeke aims to build a fully synthetic genome of S. cerevisiae which increases the genome stability by removing all repeated sequences (tRNA, transposable elements, etc.), and implements SCRaMbLE (for Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution), an inducible, high-throughput chromosome rearrangement system. This design is highly conservative with respect to gene content, the deletion of several classes of repeated sequences and the introduction of thousands of designer changes. However, it may affect genome organization and potentially alter cellular functions. To determine wether those modifications affected the three-dimensional conformation of synthetic chromosmes, we investigated it using chromosomes conformation capture coupled to second generation sequencing method (Hi-C). Currently, eight synthetic chromosomes (synI, synII, synIII, synV, synVI, synIX-R, synX et synXII) have been fully assembled. Using these strains we observed that the large-scale genomic organization is globally unaffected by the presence of synthetic chromosome(s). Two exceptions are synIII, which lacks the silent mating-type cassettes, and synXII, specifically when the ribosomal DNA is moved to another chromosome. We also exploited the contact maps to detect rearrangements induced in these SCRaMbLE strains.
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Guillaume Mercy. L'organisation 3D des chromosomes synthétiques de levure. Génomique, Transcriptomique et Protéomique [q-bio.GN]. Sorbonne Université, 2018. Français. ⟨NNT : 2018SORUS034⟩. ⟨tel-02111002⟩

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