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

Author:

Forget StephanieORCID,Krawczyk MikaylaORCID,Knight AndersORCID,Ching CharleneORCID,Copeland RachelleORCID,Mahmoodi NiushaORCID,Mayo MelissaORCID,Nguyen JamesORCID,Tan AmandaORCID,Miller MattORCID,Vroom JonathanORCID,Lutz StefanORCID

Abstract

AbstractEnzymatic DNA synthesis, using stepwise nucleotide addition catalyzed by template-independent polymerases, promises higher efficiency, quality, and sustainability than today’s 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.

Publisher

Cold Spring Harbor Laboratory

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