Enhancing the reverse transcriptase function in Taq polymerase via AI-driven multiparametric rational design.

Journal: Frontiers in bioengineering and biotechnology
Published Date: Dec 10, 2024

Abstract

INTRODUCTION: Modification of natural enzymes to introduce new properties and enhance existing ones is a central challenge in bioengineering. This study is focused on the development of Taq polymerase mutants that show enhanced reverse transcriptase (RTase) activity while retaining other desirable properties such as fidelity, 5'- 3' exonuclease activity, effective deoxyuracyl incorporation, and tolerance to locked nucleic acid (LNA)-containing substrates. Our objective was to use AI-driven rational design combined with multiparametric wet-lab analysis to identify and validate Taq polymerase mutants with an optimal combination of these properties.

Authors

  • Yulia E Tomilova
    AO Vector-Best, Novosibirsk, Russia.
  • Nikolay E Russkikh
    AcademGene LLC, Novosibirsk, Russia.
  • Igor M Yi
    AcademGene LLC, Novosibirsk, Russia.
  • Elizaveta V Shaburova
    MSU Institute for Artificial Intelligence, Lomonosov Moscow State University, Moscow, Russia.
  • Viktor N Tomilov
    SibEnzyme Ltd., Novosibirsk, Russia.
  • Galina B Pyrinova
    AO Vector-Best, Novosibirsk, Russia.
  • Svetlana O Brezhneva
    AO Vector-Best, Novosibirsk, Russia.
  • Olga S Tikhonyuk
    AO Vector-Best, Novosibirsk, Russia.
  • Nadezhda S Gololobova
    AO Vector-Best, Novosibirsk, Russia.
  • Dmitriy V Popichenko
    AO Vector-Best, Novosibirsk, Russia.
  • Maxim O Arkhipov
    AO Vector-Best, Novosibirsk, Russia.
  • Leonid O Bryzgalov
    AO Vector-Best, Novosibirsk, Russia.
  • Evgeniy V Brenner
    AO Vector-Best, Novosibirsk, Russia.
  • Anastasia A Artyukh
    AO Vector-Best, Novosibirsk, Russia.
  • Dmitry N Shtokalo
    AcademGene LLC, Novosibirsk, Russia.
  • Denis V Antonets
    MSU Institute for Artificial Intelligence, Lomonosov Moscow State University, Moscow, Russia.
  • Mikhail K Ivanov
    AO Vector-Best, Novosibirsk, Russia.

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