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

Year: 2020
Type: article
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 d TPT3 –d NaM 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: 10.1021/jacs.9b09808
Authors: Michael P. Ledbetter, Jonathan M. Craig, Rebekah J. Karadeema, Matthew T. Noakes, Hwanhee C. Kim, Sarah J. Abell, Jesse R. Huang, Brooke A. Anderson, Ramanarayanan Krishnamurthy, Jens H. Gundlach, Floyd E. Romesberg
Language: English
Open Access status: green
Awards: G1041611928, G134900822, G1556647868, G307239250, G4773797742, G5045525407, G5739519380, G8354083096
OpenAlex ID: W3003234996
FWCI: 1.406
Cites: 28
Cited by: 38
Related to: 10