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Title:Single molecule visualization of a powerful genetic engineering tool "Cas9" in action
Author(s):Singh, Dingvijay
Biophysics and Computational Biology
Abstract:Cas9 is a tiny molecular machine that has paved way for a new-era of biotech revolution. The protein-Cas9, is an endonuclease, meaning that it can cleave DNA. The ability to easily program Cas9 to cleave at specific DNA sites forms the bedrock of the promise Cas9 holds for a wide variety of genetic-engineering applications. It has already been used to make genetic changes in plants & animals. Cas9 can be mixed with a special guide-RNA to form surveillance machine to hit right DNA-targets. The sequence of the guide RNA determines the specificity of the DNA-targets by virtue of base-pairing between guide-RNA & DNA-target. But Cas9 does not always hit the right targets causing off-target effects. My research involves the use of single molecule-FRET to investigate this problem, the mechanism of Cas9 action and kinetics of Cas9-binding onto a wide variety of DNA-targets. My image attempts to show the schematic of my experiments and showcase the power of science that enabled us real-time visualization of biological events at the single-molecule level. The DNA-targets are selectively immobilized on a passivated-surface with Cas9-complex in solution. These molecules are labeled with bright-dyes that enable their detection and thus their interaction-behavior can be studied in-singulo.
Issue Date:2015-04
Rights Information:Copyright 2015 Digvijay Singh
Date Available in IDEALS:2015-04-27

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