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The use of RNA affinity tags in RNA biology
Vaškovičová, Michaela ; Svoboda, Petr (advisor) ; Drbal, Karel (referee)
RNA molecules play essential role in many important processes in living organisms. Messenger RNAs transmit genetic information from the DNA to the cellular sites of protein synthesis. RNAs, which do not encode for proteins, have numerous specialized functions in regulation of gene expression, splicing, transport, localization, translation, and stability of RNAs. For example, ribosomal and transfer RNAs are required for the translation of proteins; small nuclear RNAs are involved in splicing, and small nucleolar RNAs modify other RNAs. MicroRNAs and small interfering RNAs are essential regulators of gene expression. Other non-coding RNAs regulate chromatin and transcription. RNAs are typically present in ribonucleoprotein (RNP) complexes containing various proteins, which are essential for functions of the complexes. Understanding RNA-protein interactions and composition of RNP complexes is critical for delineating their functions. The common strategy in the study of RNP complexes is tagging the RNA component, in order to isolate, purify and analyze the tagged RNAs with their binding partners. Tagged RNP complexes can be subsequently subjected to further studies. In the first part of my work, I describe four basic strategies of tagging the RNA, which can be used for study of RNAs and RNP complexes....
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