Two Classes of Small Antisense RNAs in Fungal RNA Silencing Triggered by Non-integrative Transgenes

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2003 Jul 26

PMID 12881432

Citations 59

Authors

Francisco E Nicolas

Santiago Torres-Martinez

Rosa M Ruiz-Vazquez

Affiliations

Soon will be listed here.

Abstract

Transformation of Mucor circinelloides with self-replicative plasmids containing a wild-type copy of the carotenogenic gene carB causes silencing of the carB function in 3% of transformants. Genomic analyses revealed a relationship between silenced phenotype and number of copies of plasmids. This phenotype results from a reduction of the steady-state levels of carB mRNA, a reduction that is not due to differences in the level of transcription, indicating that silencing is post-transcriptional. Small sense and antisense RNAs have been found to be associated with gene silencing in M. circinelloides. Two size classes of small antisense RNAs, differentially accumulated during the vegetative growth of silenced transformants, have been detected: a long 25-nucleotide RNA and a short 21-nucleotide RNA. Secondary sense and antisense RNAs corresponding to sequences of the endogenous gene downstream of the initial triggering molecule have also been detected, revealing the existence of spreading of RNA targeting in fungi. These findings, together with the self-replicative nature of the triggering molecules, make M. circinelloides a suitable organism for investigating some unresolved questions in RNA silencing.

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