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RNA silencing in Chlamydomonas: mechanisms and tools

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Abstract

The generation of a comprehensive EST library and the sequencing of its genome set the stage for reverse genetics approaches in Chlamydomonas reinhardtii. However, these also require tools for the specific downregulation of target gene expression. Consequently, a large number of diverse constructs were developed aimed at reducing target gene expression in Chlamydomonas via the stable expression of antisense or inverted repeat-containing RNA. Double-stranded RNA (dsRNA) generated by the annealing of antisense and sense RNAs or by hairpin formation of an inverted repeat, feeds into the RNA silencing pathway. In this pathway, dsRNA is cleaved into ~25-bp small interfering RNAs (siRNAs) by the endonuclease Dicer. One of the two complementary strands of a siRNA is then loaded onto an Argonaute-like protein present as core component within larger complexes. Guided by this single-stranded RNA, the Argonaute-like protein either detects homologous transcripts and cleaves these endonucleolytically, or initiates transcriptional gene silencing. This article summarizes current information derived mainly from the Chlamydomonas genome project on components that are assumed to be involved in RNA silencing mechanisms in Chlamydomonas. Furthermore, all approaches employed in Chlamydomonas to date to downregulate target gene expression by antisense or inverted repeat constructs are reviewed and discussed critically.

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Acknowledgements

I would like to thank Christoph Beck, Felix Willmund, Mukesh Lodha, Cuimin Liu, and Miriam Schulz-Raffelt for their critical reading of the manuscript. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (Schr 617/4-2).

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  1. Institute of Biology II/Plant Biochemistry, University of Freiburg, Schänzlestr. 1, 79104, Freiburg, Germany

    Michael Schroda

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Schroda, M. RNA silencing in Chlamydomonas: mechanisms and tools. Curr Genet 49, 69–84 (2006). https://doi.org/10.1007/s00294-005-0042-1

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