The amazing genetic scissors called CRISPR / Cas9, the discovery that won the 2020 Nobel Prize in Chemistry, are sometimes cut in places that are not designed to target. While CRISPR has completely changed the pace of fundamental research by allowing scientists to quickly edit genetic sequences, it is working so fast that it is difficult for scientists to see what is sometimes going on. wrong and find out how to improve it.
Julene Madariaga Marcos, a Humboldt graduate, and colleagues in Professor Ralf Seidel’s laboratory at the University of Leipzig in Germany, discovered a way to analyze the ultra-fast motions of CRISPR enzymes, which help researchers understand how to identify their target chains in the hope of developing distinctiveness. Madariaga Marcos will present the research on Tuesday, February 23rd at the 65th Annual Meeting of the Biophysical Society.
To use the CRISPR enzyme to edit gene sequences, scientists can tailor it to target a specific sequence within the three billion pairs of DNA bases in the human genome. During target identification the CRISPR enzyme unlocks the DNA strands to find a sequence that contributes to the linked RNA sequence of CRISPR. But sometimes the RNA matches DNA sequences that are not very supportive. To solve this unusual game, scientists need to be able to see how CRISPR works with individual DNA base pairs, but the process is quick and difficult to watch.
To measure CRISPR activities on an ultra-fast time frame, Madariaga Marcos and colleagues turned to origami DNA, which uses specific DNA sequences to create complex three-dimensional nanostructures instead of a simple double helix. DNA origami has applications in drug delivery, nanoelectronics, and even art.
Using origami DNA, they lifted rotor arms out of DNA so that they could see with a high-speed camera on a microscope without binding the DNA with the CRISPR enzyme, causing the rotor arm to spin like blades. helicopter. With this system, they were able to quantify the different responses to games and inconsistencies within the DNA sequence.
We can directly measure the energy landscape of CRISPR / Cascade when it interacts with DNA for the first time. “
Julene Madariaga Marcos, Postdoctoral Fellow, Humboldt
This device will help scientists better understand the enzyme CRISPR, and how it will eventually land. That way, they can find out how you can increase CRISPR so that it makes fewer games off target. In the future, Madariaga Marcos is interested in “developing more tools and techniques for studying these gene editing processes in new and more detailed ways.”