| Grant number: | 12/00195-3 |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| Start date: | October 01, 2012 |
| End date: | February 28, 2017 |
| Field of knowledge: | Biological Sciences - Biochemistry - Molecular Biology |
| Principal Investigator: | Katlin Brauer Massirer |
| Grantee: | Katlin Brauer Massirer |
| Host Institution: | Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil |
| City of the host institution: | Campinas |
| Associated scholarship(s): | 15/25134-5 - Implementation of CLIP-seq computational analyzes for determination of target mRNAs and Caprin-1 protein binding sites,
BP.MS 14/20174-6 - Implementation of CLIP-seq technique and evaluation of mRNAs-target by protein Caprin-1, BP.MS |
Abstract
RNA binding proteins (RBPs) are involved in a variety of post-transcriptional cellular events such as alternative splicing, pre-mRNA transport out of the nucleus, stabilization of mRNAs and the microRNA pathway. Disruption of these proteins has been linked to pathological conditions including muscular myotonic distrophy, autism and Parkinson's. Additionally many RBPs regulate the manitanance of pluripotency and the process leading to cellular differentiation. However, little is known about the function and the mRNA binding targets for most RBPs. In the current project, we propose to combine molecular techniques and bioinformatics to identify the protein complexes containing the RBPs and to identify and characterize the mRNA targets for RBPs related to stemcellness and neuronal differentiation. We hope this study will contribute to a deeper understanding of neurological diseases in general. We propose to use immunoprecipitation combined with mass spectrometry to identify RNA binding complexes, by overexpressing RBPs in HEK293 cells. At the same time, human embryonic stem cells will be subjected to the CLIP-seq technique (cross-linking immunoprecipitation followed by next-generation sequencing), to define the profile of mRNA targets for RBPs and to map the target binding sites. Discovered post-transcriptional events will be validated in human embryonic stem cell compared to neurons derived from these cells. (AU)
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