4.5 Article

CRISPR/Cas9-mediated genome editing assists protein dynamics studies in live cells

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EUROPEAN JOURNAL OF CELL BIOLOGY
卷 101, 期 2, 页码 -

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ELSEVIER GMBH
DOI: 10.1016/j.ejcb.2022.151203

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CRISPR; Cas9 genome editing; Molecular dynamics; Fluorescence microscopy

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Studying molecular dynamics is crucial in understanding cell fate and the perturbations causing disease. Genome editing protocols now allow us to study molecular dynamics while avoiding issues associated with overexpression.
Spatial and temporal regulation of molecular reactions dictates cell fate. Thus, studying molecular dynamics is essential to understand how cells decide what to do and the fundamental perturbations causing disease. Classically, molecular dynamics has been studied by protocols based in the overexpression of fluorescent fusion proteins. However, overexpression is associated to altered stoichiometry, molecular dynamics and subcellular distribution. We here discuss the necessity to study molecular dynamics of fluorescent fusion proteins expressed under physiological mechanisms in the cell, pointing to CRISPR/Cas9-mediated genome editing as the ideal means to do so. Current genome editing protocols enable us to study molecular dynamics while avoiding drawbacks associated to overexpression.

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