Utveckling av nya sekvensbaserade strategier för att karaktärisera RNA och post-transkriptionell reglering in humana vävnadsprover
Tidsperiod: 2013-01-01 till 2015-12-31
Projektledare: Lars Feuk
Budget: 2 400 000 SEK
Over the past few years, new sequencing technologies have led to the discovery of novel types of RNA and novel catalytic functions for RNA in the cell. RNA-sequencing has further increased the ability to perform unbiased screening of RNA, expression levels and alternative splicing. We have developed a new protocol for extracting the RNA from the nucleus and the cytosol separately from tissue samples, and preliminary data indicate that this gives us a higher resolution to characterize RNA and splicing kinetics. Based on RNA sequencing and our new protocol of RNA extraction, we aim to investigate the dynamics of splicing and to identify which introns are spliced co-transcriptionally and which are spliced post transcription. We will further characterize RNA across the entire size spectrum, ranging from microRNA to long pre-mRNA in several different adult and fetal human tissues. Finally, we will also sequence the DNA of these samples to enable identification of sites that undergo RNA editing. These sequence based strategies will make it possible to answer some long-standing questions about transcription and splicing that have not been possible to address previously.