Understanding the Photophysics of the Spinach鈥揇FHBI RNA Aptamer鈥揊luorogen Complex To Improve Live-Cell RNA Imaging

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• 作者：Kyu Young Han ; Benjamin J. Leslie ; Jingyi Fei ; Jichuan Zhang ; Taekjip Ha
• 刊名：Journal of the American Chemical Society
• 出版年：2013
• 出版时间：December 18, 2013
• 年：2013
• 卷：135
• 期：50
• 页码：19033-19038
• 全文大小：426K
• 年卷期：v.135,no.50(December 18, 2013)
• ISSN：1520-5126

文摘

The use of aptamer鈥揻luorogen complexes is an emerging strategy for RNA imaging. Despite its promise for cellular imaging and sensing, the low fluorescence intensity of the Spinach鈥揇FHBI RNA aptamer鈥揻luorogen complex hampers its utility in quantitative live-cell and high-resolution imaging applications. Here we report that illumination of the Spinach鈥揻luorogen complex induces photoconversion and subsequently fluorogen dissociation, leading to fast fluorescence decay and fluorogen-concentration-dependent recovery. The fluorescence lifetime of Spinach鈥揇FHBI is 4.0 卤 0.1 ns irrespective of the extent of photoconversion. We detail a low-repetition-rate illumination scheme that enables us to maximize the potential of the Spinach鈥揇FHBI RNA imaging tag in living cells.