Discovering miRNAs: Ph.D. thesis

Søren Peter Jonstrup

Research output: Book/ReportCompendium/lecture notes

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Abstract

MicroRNAs (miRNAs) are a class of non-coding RNAs that act as negative regulators of gene expression. Here I present the identification of six new miRNAs. Furthermore, the involvement of miRNAs in osteoblast differentiation is investigated. Artificial regulation of miR-34a, miR-92, and miR-210 is shown to affect osteoblast differentiation of mesenchymal stem cells (MSCs). However, no miRNAs showing an expression pattern dependent on this differentiation was found. In stead four miRNAs (miR-7, miR-26b, miR-34a, and miR-210) are identified which expression pattern shows dependency on cell density. This cell density dependent expression is found to be conserved in several human cell lines. I also present data indicating that miR-17-5p and miR-20a are involved in both proper spermatogenesis as well as development of testis cancer. I played a large role in implementation of the microarray technique in our laboratory. I also developed an experimental approach for identification of miRNA targets capable of selecting mRNAs showing complementarity towards a specific miRNA. In a test of LNA knockdown probes I found that probes based on deoxyribose nucleotides or on 2’-O-methyl modified nucleotides works better than probes based on ribose nucleotides. Finally, a miRNA detection method relying on padlock probes is presented. This method is shown to discriminate between closely related miRNAs and only require few nanograms of total RNA for detection.
Original languageEnglish
Place of PublicationAarhus university
Number of pages135
Publication statusPublished - 2008
Externally publishedYes

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