TY - JOUR
T1 - Synthetic evolutionary origin of a proofreading reverse transcriptase
AU - Ellefson, Jared W.
AU - Gollihar, Jimmy
AU - Shroff, Raghav
AU - Shivram, Haridha
AU - Iyer, Vishwanath R.
AU - Ellington, Andrew D.
PY - 2016/6/24
Y1 - 2016/6/24
N2 - 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.
AB - 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.
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U2 - 10.1126/science.aaf5409
DO - 10.1126/science.aaf5409
M3 - Article
C2 - 27339990
AN - SCOPUS:84976337073
SN - 0036-8075
VL - 352
SP - 1590
EP - 1593
JO - Science
JF - Science
IS - 6293
ER -