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© Research
Publication : Fungal genetics and biology : FG & B

Developing a tetO/TetR system in Neurospora crassa

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Fungal genetics and biology : FG & B - 09 Dec 2019

Nguyen TS, Gladyshev E

Link to Pubmed [PMID] – 31821884

Fungal Genet. Biol. 2019 Dec;136:103316

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.

https://www.ncbi.nlm.nih.gov/pubmed/31821884