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The effects of long double-stranded RNA expression in mammalian cells.
Nejepínská, Jana ; Svoboda, Petr (advisor) ; Petr, Jaroslav (referee) ; Štefl, Richard (referee)
Double stranded RNA (dsRNA) is a foreign molecule that arises in the cell either as a by-product of viral replication or it is produced by the intramolecular or intermolecular pairing of complementary RNAs, often originating from repetitive sequences. In mammals, dsRNA can enter one of three pathways: the sequence-specific RNA silencing, the sequence-independent interferon (IFN) response, or editing by adenosine deaminases. The main focus of my PhD project was to comprehensively analyze the effects of the expressed dsRNA in mammals in the context of the whole organism. To follow this aim, we generated a construct expressing dsRNA in a form of an mRNA containing a long perfect hairpin structure. Transgenic mice ubiquitously expressing dsRNA were viable and, in contrast to the previous studies, the IFN response was not activated. In somatic cells, dsRNA was poorly processed into small interfering RNAs, did not cause transcriptional silencing in trans, and underwent low adenosine deamination without the nuclear retention. Consistent results were obtained in human cells transiently transfected with a dsRNA-expressing plasmid. On the other hand, dsRNA expression caused robust RNA interference (RNAi) in oocytes. Thus, we show for the first time that expressed dsRNA, in contrast to many other forms of...
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The effects of long double-stranded RNA expression in mammalian cells.
Nejepínská, Jana ; Svoboda, Petr (advisor) ; Petr, Jaroslav (referee) ; Štefl, Richard (referee)
Double stranded RNA (dsRNA) is a foreign molecule that arises in the cell either as a by-product of viral replication or it is produced by the intramolecular or intermolecular pairing of complementary RNAs, often originating from repetitive sequences. In mammals, dsRNA can enter one of three pathways: the sequence-specific RNA silencing, the sequence-independent interferon (IFN) response, or editing by adenosine deaminases. The main focus of my PhD project was to comprehensively analyze the effects of the expressed dsRNA in mammals in the context of the whole organism. To follow this aim, we generated a construct expressing dsRNA in a form of an mRNA containing a long perfect hairpin structure. Transgenic mice ubiquitously expressing dsRNA were viable and, in contrast to the previous studies, the IFN response was not activated. In somatic cells, dsRNA was poorly processed into small interfering RNAs, did not cause transcriptional silencing in trans, and underwent low adenosine deamination without the nuclear retention. Consistent results were obtained in human cells transiently transfected with a dsRNA-expressing plasmid. On the other hand, dsRNA expression caused robust RNA interference (RNAi) in oocytes. Thus, we show for the first time that expressed dsRNA, in contrast to many other forms of...
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