Synthetic evolutionary origin of a proofreading reverse transcriptase

Jared W. Ellefson, Jimmy Gollihar, Raghav Shroff, Haridha Shivram, Vishwanath R. Iyer, Andrew D. Ellington

Research output: Contribution to journalArticlepeer-review

67 Scopus citations

Abstract

Most reverse transcriptase (RT) enzymes belong to a single protein family of ancient evolutionary origin. These polymerases are inherently error prone, owing to their lack of a proofreading (3- 5 exonuclease) domain. To determine if the lack of proofreading is a historical coincidence or a functional limitation of reverse transcription, we attempted to evolve a high-fidelity, thermostable DNA polymerase to use RNA templates efficiently. The evolutionarily distinct reverse transcription xenopolymerase (RTX) actively proofreads on DNA and RNA templates, which greatly improves RT fidelity. In addition, RTX enables applications such as single-enzyme reverse transcription-polymerase chain reaction and direct RNA sequencing without complementary DNA isolation. The creation of RTX confirms that proofreading is compatible with reverse transcription.

Original languageEnglish (US)
Pages (from-to)1590-1593
Number of pages4
JournalScience
Volume352
Issue number6293
DOIs
StatePublished - Jun 24 2016

ASJC Scopus subject areas

  • General

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