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Analysis of transcription factor and histone modification dynamics in the nucleus of single living cells using a novel antibody-based imaging approach

Abstract : In eukaryotic cells, gene transcription is controlled by a plethora of protein complexes. However, most of our basic knowledge about transcription regulation originate from biochemical experiments or immunofluorescence experiments using fixed cells. Consequently, many efforts have been devoted recently to obtain information about the dynamic movements or assembly of transcription factors directly from living cells. Therefore, we developed a labeling strategy, named versatile antibody-based imaging approach (VANIMA), in which fluorescently labeled antibodies are introduced into living cells to image specific endogenous proteins or posttranslational modifications. We were able to show that VANIMA can be used to study dynamical processes of fundamental biological mechanisms including factors of the transcription machinery as well as histone modifications in living human cancer cells using conventional or super-resolution microscopy. Hence, in the future VANIMA will serve as a valuable tool to uncover the dynamics of endogenous biological processes including transcription directly in single living cells.
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Sascha Conic. Analysis of transcription factor and histone modification dynamics in the nucleus of single living cells using a novel antibody-based imaging approach. Genomics [q-bio.GN]. Université de Strasbourg, 2018. English. ⟨NNT : 2018STRAJ081⟩. ⟨tel-02917971⟩

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