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Volume 151, Issue 11

Knockdown of in the mushroom by double-stranded RNA-mediated gene silencing Free

Abstract

The basidiomycete has many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of the dsRNA expression construct (dsRNA) resulted in genomic insertion of dsRNA and paucity of the transcript. First, dsRNA was transformed into the homothallic strain to generate a homozygote in which both nuclei had a copy of dsRNA. The -repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy of dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach in research.

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Keyword(s): DAPI, 4′,6-diamidino-2-phenylindole , PAB, 4-aminobenzoic acid , RNAi, RNA interference and SC, synaptonemal complex
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2005-11-01
2024-04-19
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Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA-mediated gene silencing
Microbiology 151, 3669 (2005); https://doi.org/10.1099/mic.0.28209-0
/content/journal/micro/10.1099/mic.0.28209-0
/content/journal/micro/10.1099/mic.0.28209-0
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