Transcription factors dynamics in stem cells using fluorescence correlation spectroscopy (FCS).

ANGIOLINI J; OSES C; LEVI V; VERNERI P; GUBERMAN A

Congreso; II LAFeBS, IX IberoAmerican Congress of Biophysics, XLV Reunion Anual SAB 2016; 2016

Embryonic stem cells (ESC) can differentiate to cells derived from allthree germ layers and have an unlimited possibility of self renewal.These properties determine the relevance of ESC studies for both, understandingembryo development and applications inregenerative medicine. Pluripotency depends on specific transcriptionfactors (TFs), such as Oct4, Sox2 and Nanog. They induce genes necessaryto preserve undifferentiated state and repress genes related todifferentiation.The transcription machinery and regulatory components are recruiteddynamically on their target genes. This means that the dynamics ofinteraction plays a fundamental role in the different stages involved ingene expression. Moreover, these interactions are modulated by thelocal plasticity of the chromatin.In this work we explore the dynamics of TFs relevant for pluripotency inESC.With this aim we performed fluorescence correlation spectroscopy(FCS) in ESC transfected with vectors encoding TFs fused to either enhancedor photoactivatable green fluorescent protein (GFP). To correlatethe dynamics of the TFs with the chromatin organization, we cotransfectedthe cells with chromatin associated proteins fused to red fluorescentproteins.