Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair

Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair

Unnatural base pairs (UBPs) have been developed and used for a variety of in vitro applications as well as for the engineering of semisynthetic organisms (SSOs) that store and retrieve increased information. However, these applications are limited by the availability of methods to rapidly and accura...

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Veröffentlicht in:Journal of the American Chemical Society 2020-02, Vol.142 (5), p.2110-2114
Hauptverfasser: Ledbetter, Michael P, Craig, Jonathan M, Karadeema, Rebekah J, Noakes, Matthew T, Kim, Hwanhee C, Abell, Sarah J, Huang, Jesse R, Anderson, Brooke A, Krishnamurthy, Ramanarayanan, Gundlach, Jens H, Romesberg, Floyd E
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Base Pairing
Genetic Code
Hydrophobic and Hydrophilic Interactions
Nanopores
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container_end_page 2114
container_issue 5
container_start_page 2110
container_title Journal of the American Chemical Society
container_volume 142
creator Ledbetter, Michael P
Craig, Jonathan M
Karadeema, Rebekah J
Noakes, Matthew T
Kim, Hwanhee C
Abell, Sarah J
Huang, Jesse R
Anderson, Brooke A
Krishnamurthy, Ramanarayanan
Gundlach, Jens H
Romesberg, Floyd E
description Unnatural base pairs (UBPs) have been developed and used for a variety of in vitro applications as well as for the engineering of semisynthetic organisms (SSOs) that store and retrieve increased information. However, these applications are limited by the availability of methods to rapidly and accurately determine the sequence of unnatural DNA. Here we report the development and application of the MspA nanopore to sequence DNA containing the dTPT3–dNaM UBP. Analysis of two sequence contexts reveals that DNA containing the UBP is replicated with an efficiency and fidelity similar to that of natural DNA and sufficient for use as the basis of an SSO that produces proteins with noncanonical amino acids.
doi_str_mv 10.1021/jacs.9b09808
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source ACS Publications; MEDLINE
subjects Base Pairing
Genetic Code
Hydrophobic and Hydrophilic Interactions
Nanopores
title Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair
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