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Developing a tetO/TetR system in Neurospora crassa

Abstract : The development of a tetO/TetR system in the fungus Neurospora crassa is described. The system includes (i) a synthetic gene encoding a TetR variant fused to GFP, and (ii) a standard tetO array integrated homologously, as a proof of principle, near the his-3 gene. The localization of TetR-GFP at the tetO array (observed by fluorescence microscopy) can be disrupted by the application of tetracycline. The full-length array is stable during vegetative growth, but it triggers strong repeat-induced point mutation (RIP) by the RID-dependent as well as the DIM-2-dependent pathways during the sexual phase. Thus, both RIP pathways must be inactivated to allow the faithful inheritance of the unmodified construct. In summary, this study introduces a new molecular tool into Neurospora research, and suggests that the standard tetO array can self-engage in recombination-independent homologous pairing.
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Submitted on : Thursday, July 21, 2022 - 9:10:40 AM
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Tinh-Suong Nguyen, Eugene Gladyshev. Developing a tetO/TetR system in Neurospora crassa. Fungal Genetics and Biology, Elsevier, 2020, 136, pp.103316. ⟨10.1016/j.fgb.2019.103316⟩. ⟨pasteur-03261456⟩



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