"A novel look at the intermediate filament vimentin through new CRISPR technology "
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2018-05
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Knudtson, Katie
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Essner, Jeffrey
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Abstract
The cytoskeleton is required to maintain a cell’s shape and mediate cellular signaling and movement. Intermediate filaments are a component of the cytoskeleton, and vimentin is an intermediate filament associated with endothelium and mesenchymal tissue (Evans, 1998). Because of vimentin’s intimate relationship with cellular movement, it is not surprising that it has been associated with many metastatic cancers. A few past studies provide some evidence that loss of vimentin in mice produces a vascular phenotype (Eckes et al. 2000; Eckes et al., 1998; Nieminen et al., 2006; Schiffers et al., 2000). Previous work in our lab has shown that a transient knockdown of vimentin in zebrafish produces misdirection of developing vasculature. In this study, we utilize a novel CRISPR system to extend these experiments by creating a knockout of the vimentin gene, serving to validate the knockdown phenotype and determine whether vimentin is developmentally necessary. We also create a knockin fusion protein with red fluorescent protein to examine cellular localization during vascular morphogenesis. Our fusion protein knockin shows that vimentin is present in the nervous system 3 days post fertilization (dpf). Preliminary data from knockout mutations supports a role for vimentin during vascular morphogenesis, including migration, branching, and lumen formation.
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