Selection of Intracellularly Functional RNA Mimics of Green Fluorescent Protein Using Fluorescence-Activated Cell Sorting

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• 作者：Jiawei Zou ; Xin Huang ; Lei Wu ; Gangyi Chen ; Juan Dong…
• 关键词：In vivo ; FACS ; Fluorescence ; Aptamers
• 刊名：Journal of Molecular Evolution
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：81
• 期：5-6
• 页码：172-178
• 全文大小：1,279 KB
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• 作者单位：Jiawei Zou (1)
  Xin Huang (1)
  Lei Wu (1)
  Gangyi Chen (1)
  Juan Dong (1)
  Xin Cui (1)
  Zhuo Tang (1)
  
  1. Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People鈥檚 Republic of China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Cell Biology
  Microbiology
  Plant Sciences
  
• 出版者：Springer New York
• ISSN：1432-1432

文摘

Fluorescence-activated cell sorting (FACS) was exploited to isolate Escherichia coli cells that were highly fluorescent due to the expression of RNA aptamers that induce fluorescence of 3,5-difluoro-4-hydroxybenzylidene imidazolinone. Two different aptamers, named ZT-26 and ZT-324, were identified by this method and compared to the fluorescence-signaling properties of Spinach, a previously reported RNA aptamer. Aptamer ZT-26 exhibits significantly enhanced fluorescence over Spinach only in vitro. However, aptamer ZT-324 is 36 % brighter than Spinach when expressed in E. coli. The FACS-based selection strategy presented here is attractive for deriving fluorescent RNA aptamers that function in cells as it directly selects for cells with a high level of fluorescence due to the expression of the RNA aptamer. Keywords In vivo FACS Fluorescence Aptamers