Expression analysis of Drosophila melanogaster microRNAs
Başlık çevirisi mevcut değil.
- Tez No: 401189
- Danışmanlar: PROF. DR. THOMAS TUSCHL
- Tez Türü: Doktora
- Konular: Tıbbi Biyoloji, Medical Biology
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2006
- Dil: İngilizce
- Üniversite: Georg-August-Universität Göttingen
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 148
Özet
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Özet (Çeviri)
miRNAs are tiny regulatory RNAs of 20-25 nucleotides (nt) in length that are endogenously expressed in cells. Hundreds of miRNA genes have been identified in plants and animals. Together they form a large family of regulatory molecules that control important cellular and developmental pathways. Most animal miRNAs regulate the translation of different cellular mRNAs via imperfect base pairing interactions. Due to the complexity of target mRNA recognition only a small subset of miRNAs is annotated with biological functions. In order to contribute to their functional identification, I identified the temporal and spatial expression pattern of D. melanogaster miRNAs by Northern blot analysis at different developmental stages. Temporal expression patterns of miRNAs can be used for the verification of predicted miRNA targets and biological functions. Tissue specific expression patterns of M. musculus miRNAs were identified again by northern blot analysis. Several miRNAs are highly tissue specifically expressed in adult mouse suggesting that they may have important tissue specific biological functions. For the identification of spatial expression patterns of D. melanogaster miRNAs, I tried to optimize the whole-mount in situ hybridization protocol for fly embryos. Our efforts to identify an efficient method for the identification of spatial expression patterns of D. melanogaster miRNAs were not successful. Detecting fly miRNAs by in situ hybridization is challenging and only few miRNAs were detected so far. I targeted miRNA processing factors by RNAi in order to determine the effects on miRNA biosynthesis in D. melanogaster cells. We identified dmDGCR8 (Pasha), which is a Drosha interacting protein, as a novel factor involved in nuclear processing of pri-miRNAs. I demonstrated that dmDGCR8 and Drosha depletions resulted in primiRNA accumulation in the cells. In order to completely deplete Drosha from the embryos I tried to knock out the Drosha gene with the P-element mobilization method. I could not delete Drosha gene by this method.
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