Genetic engineering mechanism visualized - School of Science, the University of Tokyo
Nov 16, 2017

Genetic engineering mechanism visualized


Researchers at Kanazawa University and the University of Tokyo report in Nature Communications the visualization of the dynamics of 'molecular scissors' -- the main mechanism of the CRISPR-Cas9 genetic-engineering technique.


Overview of the press release

One of the techniques used in genetic engineering -- the process of artificially modifying the genome of a living organism -- involves the so-called CRISPR-Cas9 nuclease system. Using this system, a cell's DNA can be cut at a desired site, where genes can be deleted or added. Selection of the site to be cut is done by a 'guide RNA' molecule bound to the Cas9 protein. Now, a team of researchers led by Mikihiro Shibata from Kanazawa University and Osamu Nureki from the University of Tokyo has visualized the dynamics of the CRISPR-Cas9 complex, in particular how it cuts DNA, providing valuable insights into the CRISPR-Cas9-mediated DNA cleavage mechanism.

For their visualization studies, the scientists used high-speed atomic-force microscopy (HS-AFM), a method for imaging surfaces. A surface is probed by moving a tiny cantilever over it; the force experienced by the probe can be converted into a height measure. A scan of the whole surface then results in a height map of the sample. The high-speed experimental set-up of Shibata and colleagues enabled extremely fast, repeated scans -- convertible into movies -- of the biomolecules taking part in the molecular scissoring action.

Figure:HS-AFM movie of DNA cleavage by CRISPR-Cas9
The HNH nuclease domain in an active conformation is indicated by a magenta arrow.



For more information, please visit the website of Kanazawa University.

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