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Post-transcriptional regulatory networks mediated by RNA binding proteins in human pluripotent stem cells


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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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ALVES BARBOSA, EVERTON DE ALMEIDA; SERAPHIM, THIAGO VARGAS; GANDIN, CESAR AUGUSTO; TEIXEIRA, LEILANE FERREIRA; GONCALVES DA SILVA, RONNI ANDERSON; RIGHETTO, GERMANNA L.; GONCALVES, KALIANDRA DE ALMEIDA; VASCONCELLOS, RAPHAEL DE SOUZA; ALMEIDA, MARCIA ROGERIA; SILVA JUNIOR, ABELARDO; et al. Insights into the full-length SRPK2 structure and its hydrodynamic behavior. International Journal of Biological Macromolecules, v. 137, p. 205-214, . (14/07206-6, 13/50724-5, 11/23110-0, 12/00195-3, 17/03489-1, 12/50161-8, 17/07335-9)
SATO, JOAO RICARDO; VIDAL, MACIEL CALEBE; SANTOS, SUZANA DE SIQUEIRA; MASSIRER, KATLIN BRAUER; FUJITA, ANDRE. Complex Network Measures in Autism Spectrum Disorders. IEEE-ACM TRANSACTIONS ON COMPUTATIONAL BIOLOGY AND BIOINFORMATICS, v. 15, n. 2, p. 581-587, . (12/00195-3, 13/00506-1, 13/03447-6, 14/09576-5, 13/10498-6)
DE-SOUZA, EVANDRO A.; CAMARA, HENRIQUE; SALGUEIRO, WILLIAN G.; MORO, RAISSA P.; KNITTEL, THIAGO L.; TONON, GUILHERME; PINTO, SILAS; PINCA, ANA PAULA F.; ANTEBI, ADAM; PASQUINELLI, AMY E.; et al. RNA interference may result in unexpected phenotypes in Caenorhabditis elegans. Nucleic Acids Research, v. 47, n. 8, p. 3957-3969, . (12/00195-3, 10/52557-0, 17/22057-5, 17/04377-2, 17/03423-0, 16/02207-0, 17/08829-5, 15/01316-7, 17/01339-2, 14/10814-8, 17/01184-9, 14/25068-0, 16/15958-3)

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