Aarhus University Seal / Aarhus Universitets segl

Towards an understanding of miRNA regulation

Research output: Book/anthology/dissertation/reportPh.D. thesisResearch

miRNAs are well-known regulators of gene expression. They function post-transcriptionally by binding to complementary sites within the 3´UTR of target mRNAs, which mediates translational repression and destabilization. However, miRNA expression itself is also subjected to regulation.
Here, we report a new method to investigate and potentially characterize the pri-miRNA transcript.
Overexpression of a transdominant Drosha mutant, which is unable to cleave its substrate, enables stabilization of the pri-miRNA transcript. Drosha mutant immunoprecipitation from the nuclear compartment is performed followed by high-throughput sequencing (nuclear Drosha Mt2 RIPseq). This method allows for the detection of global pri-miRNA signature and also provides a method to potentially identify new Drosha substrates.
Furthermore, data on the identification of a novel endogenous circular RNA sponge (ciRS-7) are presented. This study discovered a non-coding circular RNA containing ~70 miR-7 binding sites that can sequester miR-7 from its mRNA targets. This functionally characterizes an endogenous expressed circular RNA for the first time.
Finally, identification of ciRS-7-associated proteins and ciRS-7 localization and transportation within the cell was pursued by investigating the use of a MS2 system to accommodate both project aims. We inserted MS2 stem-loops into ciRS-7 and preliminary results of ciRS-7 immuno-precipitation and ciRS-7 localization in the cell are presented.
Original languageEnglish
PublisherAarhus University, Science and Technology
Number of pages173
Publication statusPublished - 1 Jun 2016

See relations at Aarhus University Citationformats

ID: 85273238