Induction of gene silencing by hairpin RNA expression in Tetrahymena thermophila reveals a second small RNA pathway.
Mol Cell Biol
; 26(23): 8731-42, 2006 Dec.
Article
en En
| MEDLINE
| ID: mdl-17000759
ABSTRACT
Unlike in other eukaryotes, in which it causes gene silencing, RNA interference (RNAi) has been linked to programmed DNA deletion in the ciliate Tetrahymena thermophila. Here we have developed an efficient method to inducibly express double-stranded RNA hairpins and demonstrated that they cause gene silencing through targeted mRNA degradation in all phases of the life cycle, including growth, starvation, and mating. This technique offers a new tool for gene silencing in this model organism. Induction of RNA hairpins causes dramatic upregulation of Dicer and Argonaute family genes, revealing a system capable of rapidly responding to double-stranded RNA. These hairpins are processed into 23- to 24-nucleotide (nt) small RNAs, which are distinctly different from the 28- to 30-nt small RNAs known to be associated with DNA deletion. Thus, two different small RNA pathways appear to be responsible for gene silencing and DNA deletion. Surprisingly, expression of the RNA hairpin also causes targeted DNA deletion during conjugation, although at low efficiencies, which suggests a possible crossover of these two molecular paths.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ARN
/
Tetrahymena thermophila
/
Silenciador del Gen
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MicroARNs
Límite:
Animals
Idioma:
En
Año:
2006
Tipo del documento:
Article