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Download or read book DNA Damage Response in Live Yeast Using Single-molecule Microscopy written by Shubhika Ranjan and published by . This book was released on 2021 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: "The genetic information stored in DNA must be faithfully copied and transmitted to the next generation of cells at every cell cycle. The replication of this genetic information is performed by special multiprotein replication machinery, referred to as "replisome," which synthesizes both daughter duplexes simultaneously. At times, the accurate replication of the genome can go awry, causing mutations that can lead to a collection of diseases. In eukaryotic cells, detection and response to DNA damage during DNA replication is performed by the DNA damage response (DDR) pathway. DDR uses exposed single-stranded DNA as a signal for DNA damage and proceeds by activation of kinases that transmit the signal and change the cell program to respond to the damage. The overall purpose of this work is to understand how cells do this initial detection and how they make the decision to activate the signalling pathway. I used budding yeast (Saccharomyces cerevisiae), a unicellular model organism, to understand the eukaryotic genetic architecture because it provides a framework to develop and optimize methods to standardize the analysis. I focus on the study of dynamics of DNA replication and DDR proteins of cells experiencing DNA damage or replication fork stalling using single-molecule microscopy. Although this technique provides a high signal-to-noise ratio for visualization while still retaining the integral features in the physiological context of biological systems, various factors play a major role in attaining such high-quality data for further analysis. In the first part of this work, I provide an overview for optimizing the single-molecule techniques while considering various factors involved. In the second section, I describe the initial work towards visualizing the proteins involved in DNA damage response in HaloTag labelled S. cerevisiae"--