Working with Personal Genomics, Inc., the Taiwan Protein Project helped to demonstrate the feasibility of a new method for next generation DNA sequencing.
Taiwan Protein Project (TPP), directed by Dr Ming-Daw Tsai, is a program supported by the Executive Yuan’s policy quota. In addition to performing frontier research, an important mission of TPP is to assist domestic industry to solve protein-related problems.
Current methods commonly used in DNA sequencing poses limitations to the process. These limitations include the need to pause for cleavage at each cycle, the residual fluorescent label at the nucleobase not cleaved completely in each cycle, and the relatively short read length. The research team highlights that despite the development of many highly efficient DNA sequencing methods over the past four decades, there is room and need for further development and improvement.
The research team first showed that 3’-esterified dNTP can be incorporated into a template-primer DNA by Thermococcus sp. 9°N DNA polymerase and solved the crystal structures of the reaction intermediates and products. Then they showed that the reaction can occur continuously. Finally, they used 5’-FAM-labeled primer and esterified dNTP with a dye to show that the reaction can proceed to ca. 450 base pairs, and that the intermediates of many individual steps can be identified.
Dr Tsai said, “The results demonstrate the feasibility of a 3’-editing based DNA sequencing method that could find practical applications after further optimization.”
This cooperative research involved Dr Ming-Daw Tsai and other TPP members including Dr Wen-Jin Winston Wu and Dr Manuel Maestre-Reyna, along with the PGI team including Dr Shiuan-Woei Lin Wu (the first author), Dr Chung-Fan Chiou, and its board director and CEO Dr Johnsee Lee. The work has been published recently in Nature Research-Communications Biology on June 20, 2019.
The team has worked closely for nearly three years to lead to the present publication. A new method for DNA sequencing is also likely to benefit the development of disease diagnosis, and drug development. [APBN]