Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair,Journal of the American Chemical Society

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Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-01-27 , DOI: 10.1021/jacs.9b09808
Michael P Ledbetter ₁ , Jonathan M Craig ₂ , Rebekah J Karadeema ₁ , Matthew T Noakes ₂ , Hwanhee C Kim ₂ , Sarah J Abell ₂ , Jesse R Huang ₂ , Brooke A Anderson ₁ , Ramanarayanan Krishnamurthy ₁ , Jens H Gundlach ₂ , Floyd E Romesberg ₁

Affiliation

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.

中文翻译：

扩展遗传字母表的纳米孔测序揭示了主要疏水性非自然碱基对的高保真复制

非自然碱基对 (UBP) 已被开发并用于各种体外应用以及用于存储和检索更多信息的半合成生物 (SSO) 的工程设计。然而，这些应用受到快速准确确定非天然 DNA 序列的方法的可用性的限制。在这里，我们报告了 MspA 纳米孔的开发和应用，用于对含有 dTPT3-dNaM UBP 的 DNA 进行测序。对两个序列上下文的分析表明，包含 UBP 的 DNA 以与天然 DNA 相似的效率和保真度复制，足以用作 SSO 的基础，产生具有非规范氨基酸的蛋白质。

更新日期：2020-01-27

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