Resolving the Sequence of RNA Strands by Tip-Enhanced Raman Spectroscopy
Resolving the Sequence of RNA Strands by Tip-Enhanced Raman Spectroscopy
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DOI：
10.1021/acsphotonics.0c01486
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发表时间：
2020-12
期刊：
影响因子：
7
通讯作者：
Zhe He;Weiwei Qiu;Megan E. Kizer;Jizhou Wang;Wencong Chen;A. Sokolov;Xing Wang;Jonathan Hu;M. Scully
中科院分区：
文献类型：
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作者：
Zhe He;Weiwei Qiu;Megan E. Kizer;Jizhou Wang;Wencong Chen;A. Sokolov;Xing Wang;Jonathan Hu;M. Scully
RNA在指导蛋白质表达和催化生物反应中发挥着关键作用。金标准 RNA 测序方法需要将 RNA 转化为互补 DNA (cDNA)。随后通过聚合酶链式反应 (PCR) 和测序进行 DNA 扩增，这使得 RNA 测序变得间接、复杂且容易受到序列数据偏差的影响。本文演示了单碱基水平的 RNA 成像，同时说明了通过尖端增强拉曼散射 (TERS) 技术读取 RNA 序列的直接方法。为了解析 RNA 链内的核苷酸，我们采用了包含金尖端和金底物的间隙模式 TERS。在根据参考序列分析 TERS 测量结果后，我们以 90% 的准确度识别了 RNA 序列。这种原理验证的 RNA 成像方法显着推进了直接 RNA 测序技术，无需 RNA 标记或逆转录酶 RT-PCR 扩增。
RNA plays critical roles in guiding protein expression and catalyzing biological reactions. The gold standard RNA sequencing method requires converting RNA to complementary DNA (cDNA). This is followed by DNA amplification via polymerase chain reaction (PCR) and sequencing, making RNA sequencing indirect, complicated, and susceptible to sequence data bias. This paper demonstrates RNA imaging at the single-base level while illustrating a direct method to read RNA sequences by tip-enhanced Raman scattering (TERS) technique. To resolve nucleotides within an RNA strand, we adopted gap-mode TERS involving a gold tip and a gold substrate. After analyzing TERS measurements based on the reference sequence, we identified RNA sequences with 90% accuracy. This proof-of-principle RNA imaging method significantly advances a direct RNA sequencing technique without RNA labeling or reverse transcriptase RT-PCR amplification.