Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair
- Michael P. LedbetterMichael P. LedbetterDepartment of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United StatesMore by Michael P. Ledbetter,
- Jonathan M. CraigJonathan M. CraigDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Jonathan M. Craig,
- Rebekah J. KaradeemaRebekah J. KaradeemaDepartment of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United StatesMore by Rebekah J. Karadeema,
- Matthew T. NoakesMatthew T. NoakesDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Matthew T. Noakes,
- Hwanhee C. KimHwanhee C. KimDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Hwanhee C. Kim,
- Sarah J. AbellSarah J. AbellDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Sarah J. Abell,
- Jesse R. HuangJesse R. HuangDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Jesse R. Huang,
- Brooke A. AndersonBrooke A. AndersonDepartment of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United StatesMore by Brooke A. Anderson,
- Ramanarayanan KrishnamurthyRamanarayanan KrishnamurthyDepartment of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United StatesMore by Ramanarayanan Krishnamurthy,
- Jens H. GundlachJens H. GundlachDepartment of Physics, University of Washington, Seattle, Washington 98195, United StatesMore by Jens H. Gundlach, and
- Floyd E. Romesberg*Floyd E. Romesberg*[email protected]Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United StatesMore by Floyd E. Romesberg
Abstract
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.
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