Scientists from the University of Leipzig and the University of Vilnius created a new method that can track the gene scissors of CRISPR-Cas with the finest resolution in real time. This technique can also be utilized in various CRISPR-Cas complexes or biomolecules.
During gene recognition, the DNA of the target sequence is unwound to allow base pairing with the RNA. "The central question of the project was whether the unwinding of a piece of DNA that is only 10 nanometers (nm) long can be tracked in real-time at all," said Dominik Kauert, one of the primary authors of the study.
The scientists used DNA nanotechnology to study the unwinding process. They used this technology to create a 75 nm long DNA rotor blade with a gold nanoparticle connected to one end. In the study, the unwinding of the 10 nm long and 2 nm thin DNA sequence was turned over to the gold nanoparticle's rotation along a circle with 160 nm diameter—this magnified movement could be monitored in a unique microscope setup.
This new technique can now be utilized to analyze CRISPR-Cas sequence recognition almost base pair by base pair. The data from this study can be used in future studies to allow a better selection of RNA sequences that exclusively recognize the suitable target sequence to enhance the gene scissors' precision.
(Source: Crop Biotech Update, International Service for Acquisition of Agri-Biotech Applications.