Development of DNA Sequencing Technologies Using Gating Nanopores

利用门控纳米孔开发 DNA 测序技术

• 批准号: 20200025
• 负责人: TANIGUCHI Masateru
• 金额: $ 20.05万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20200025/
• 关键词: ナノボア; ゲーティングナノボア; DNAシーケンサー; 微細加工; 単一分子計測; トンネル電流; イオン電流; ナノポア; ゲーティングナノポア; メチル化DNA; 固体ナノポア; DNAシーケンシング; 1分子科学; 生体分析; ナノ加工; 固体ナノポ; ゲーティング固体ナノポア; 金ナノ粒子; 微小電流計測; マイクロ流路

Gating nanopores are the key devices for third generation DNA sequencing technologies. These nanodevices will make sequencing kilobase length single-stranded genomic DNA or RNA or identifying individual small molecules using only electric currents and without fluorescent labels at low cost and unheard speeds. We have developed vertical and parallel gating nanopores with embedded nanogap-electrodes in a solid-state nanopore. The vertical type consists of a single nanogap electrode with the nanopore perpendicular to the surface of the silicon substrate. The parallel type consists of a single nanogap electrode with the nanopore parallel to the surface of the substrate. We synthesized vertical gating nanopores with a diameter of 30 nm using an 11-step nanofabrication process. Vertical gating nanopores can indetify a single Au nanoparticle (~ = 28 nm) passing through them by changes in the electric current flowing between the nano-electrodes. Single base molecules of DNA can be identified by changes in tunneling current between nano-electrodes using parallel gating nanopores, incorporating a microfluidic channel in to nano-fabricated mechanically controllable break junction. We found that single-molecule electrical conductance order thymine < cytosine < adenine < guanine corresponds to the highest occupied molecular orbital (HOMO) energy order.

门控纳米孔是第三代DNA测序技术的关键器件。这些纳米器件将使测序酶长度单链基因组DNA或RNA或识别单个小分子仅使用电流，而没有荧光标记，以低成本和前所未有的速度。我们已经开发了垂直和平行门控纳米孔与嵌入式纳米间隙电极在固态纳米孔。垂直型由单个纳米间隙电极组成，纳米孔垂直于硅衬底的表面。平行型由单个纳米间隙电极组成，其中纳米孔平行于基底的表面。我们使用11步纳米纤维工艺合成了直径为30 nm的垂直门控纳米孔。垂直门控纳米孔可以通过纳米电极之间流动的电流的变化来识别通过它们的单个Au纳米颗粒（~ = 28 nm）。DNA的单个碱基分子可以通过使用平行门控纳米孔的纳米电极之间的隧穿电流的变化来识别，将微流体通道并入纳米制造的机械可控的断裂结中。我们发现，单分子电导顺序胸腺嘧啶<胞嘧啶<腺嘌呤<鸟嘌呤对应的最高占据分子轨道（HOMO）的能量顺序。

项目成果

dentifying Single Nucleotides by Tunneling Current

通过隧道电流识别单核苷酸

• 发表时间: 2010
• 期刊: Nature Nanotechnology 5
• 作者: Makusu Tsutsui;Masateru Taniguchi;Kazumichi Yokota;Tomoji Kawai
• 通讯作者: Tomoji Kawai

Identifying single nucleotides by tunnelling current

• DOI: 10.1038/nnano.2010.42
• 发表时间: 2010-04-01
• 期刊: NATURE NANOTECHNOLOGY
• 影响因子: 38.3
• 作者: Tsutsui, Makusu;Taniguchi, Masateru;Kawai, Tomoji
• 通讯作者: Kawai, Tomoji

Development of Gating Nanopores for Single-Molecule Electrical Sequencing

用于单分子电测序的门控纳米孔的开发

• 发表时间: 2011
• 作者: Onodera;S.;M. Taniguchi
• 通讯作者: M. Taniguchi

ゲーティング固体ナノポアを用いた電気計測

使用门控固态纳米孔进行电学测量

• 发表时间: 2009
• 作者: K.Tsukagoshi;S.Uryu;Y.Aoyagi;谷口正輝
• 通讯作者: 谷口正輝

Identification of Single Nucleotides Using Gating Nanopores

使用门控纳米孔鉴定单核苷酸

• 发表时间: 2010
• 作者: 北島創;安部原也;伊藤誠;平岡敏洋;丸茂喜高;和田隆広;高田翔太;M. Taniguchi
• 通讯作者: M. Taniguchi