We used a new generation of CRISPR technology, Casposons to achieve copy-paste site-directed insertion by modifying the common tool plasmids of prokaryotic expression systems, adding TSD and TIR elements recognized by Casposons. Our optimized Casponson technology can achieve gene editing without causing DNA disruption and more accurately correspond to the target gene editing tool.

At first, our experiment is performed in the molecular biology laboratory of the Experimental Research center of China Academy of the Chinese Medical Sciences, a BSL-2 laboratory. The laboratory not only meets the requirements of BSL-1 laboratories but also is safer and has effective prevention and treatment measures for pathogenic factors with moderate or potential hazards, for instance, fire extinguishers, eye washers, and other facilities, as well as the corresponding clear instructions. All strains, reagents, and consumables we used in the experiment are safe, do no harm to the ecological environment, and do not cause any harm to our human body under the standard operation of our laboratory personnel.

We have already collected the figures from our experiments. Casposons is a new gene editing tools that can implement gene insertion precisely. In our project, we set up an in vitro gene editing system with the casposons. From the result, we can find out that with the casposase protein in this system, we inserted the Kanamycin gene into the pUC19 plasmid at the TSD site.

Because there are many diseases caused by gene variation, such as gene duplication, gene deletion, gene mutation, and so on, it is important to develop a gene editing tool to solve these problems at its source. Casposons is one of the tools nowadays. In the very near future, this tool will be improved and may be used in disease treatment and bring patients a better life.