Thomas Christopher A, Brinkerhoff Henry, Craig Jonathan M, Hoshika Shuichi, Mihaylova Desislava, Pfeffer Akira M, Franzi Michaela C, Abell Sarah J, Carrasco Jessica D, Gundlach Jens H, Benner Steven A, Laszlo Andrew H
Department of Physics, University of Washington, Seattle, WA, USA.
Foundation for Applied Molecular Evolution, Alachua, FL, USA.
Nat Commun. 2025 Aug 6;16(1):7240. doi: 10.1038/s41467-025-61991-9.
"ALternative Isoinformational ENgineered" (ALIEN) DNA is a biomimetic polymer composed of four entirely anthropogenic nucleotides. These alternative nucleosides form base pairs orthogonal to canonical bases and fold into the familiar B-form DNA double-helix, endowing ALIEN DNA with valuable biotechnological applications. The ability to sequence ALIEN DNA is essential for its continued development. However traditional sequencing approaches rely on chemical recognition of ACGT-DNA and cannot be easily adapted to ALIEN DNA. Here we demonstrate de novo nanopore sequencing of DNA comprised entirely of the four anthropogenic DNA bases. We show direct, label-free, single-molecule sequencing of such nucleic acids without the requirements of fluorescent labels, transliteration, amplification, or enzymatic synthesis. This paves the way for routine, accessible, and high-accuracy sequencing of DNA beyond A, C, G, and T.
“另类等信息工程化”（ALIEN）DNA是一种由四种完全人工合成的核苷酸组成的仿生聚合物。这些另类核苷形成与标准碱基正交的碱基对，并折叠成常见的B型DNA双螺旋结构，赋予了ALIEN DNA重要的生物技术应用价值。对ALIEN DNA进行测序的能力对其持续发展至关重要。然而，传统的测序方法依赖于对ACGT-DNA的化学识别，难以轻易应用于ALIEN DNA。在此，我们展示了对完全由四种人工合成DNA碱基组成的DNA进行从头纳米孔测序。我们展示了对此类核酸的直接、无标记、单分子测序，无需荧光标记、转译、扩增或酶促合成。这为超越A、C、G和T的DNA常规、可及且高精度测序铺平了道路。
