Evolving a High-Performance Terminal Deoxynucleotidyl Transferase for Enzymatic DNA Synthesis

Forget, S.; Krawczyk, M.; Knight, A. M.; Ching, C.; Copeland, R.; Mahmoodi, N.; Mayo, M.; Nguyen, J.; Tan, A.; Miller, M. G.; Vroom, J.; Lutz, S.

doi:10.1101/2024.06.24.600205

bioRxiv · 10.1101/2024.06.24.600205

Evolving a High-Performance Terminal Deoxynucleotidyl Transferase for Enzymatic DNA Synthesis

Forget, S.·Krawczyk, M.·Knight, A. M.·Ching, C.·Copeland, R.·Mahmoodi, N.·Mayo, M.·Nguyen, J.·Tan, A.·Miller, M. G.·Vroom, J.·Lutz, S.

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Abstract

Enzymatic DNA synthesis, using stepwise nucleotide addition catalyzed by template-independent polymerases, promises higher efficiency, quality, and sustainability than todays industry standard phosphoramidite-based processes. We report on the directed evolution of a terminal deoxynucleotidyl transferase that uses 3-phosphate blocked dNTPs to control the polymerization reaction and demonstrates high activity for these modified substrates and improved template promiscuity and thermostability.

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Forget, S., Krawczyk, M., Knight, A. M., Ching, C., Copeland, R., Mahmoodi, N., Mayo, M., Nguyen, J., Tan, A., Miller, M. G., Vroom, J., Lutz, S.. 2024-06-27. Evolving a High-Performance Terminal Deoxynucleotidyl Transferase for Enzymatic DNA Synthesis. https://doi.org/10.1101/2024.06.24.600205

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