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Originally published In Press as doi:10.1074/mcp.T100002-MCP200 on November 27, 2001.
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Molecular & Cellular Proteomics 1:99-103, 2002.
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

Technology

Combinatorial Selection of RNA Ligands for Complex Cellular Targets

The RNA Liagands-Based Proteomics*

Huicheng Tian{ddagger}

From the Department of Genetics, Center for Genetic and Cellular Therapies, Duke University, Durham, North Carolina 27710

This study explores the selection of high affinity RNA ligands for the complex cellular targets present in crude HeLa nuclear extract through directed evolution and deconvolution. RNA ligands for the mixed nuclear targets were selected from around 6 x 1014 RNA sequences through an iterated enrichment process. RNA ligands for various gene products of the extract were simultaneously selected and were shown to specifically interact with their target molecules. The target molecules were isolated from the nuclear extract by affinity chromatography using columns tagged with the RNA ligands, resolved on two-dimensional gels, and identified by mass spectrometry. These RNA ligands may be useful in characterizing novel functions of cellular proteins and modulating complex molecular events.

{ddagger} To whom correspondence should be addressed. Fax: 919-942-2345; E-mail: htian{at}alumnidirector.com.


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S. Fitter and R. James
Deconvolution of a Complex Target Using DNA Aptamers
J. Biol. Chem., October 7, 2005; 280(40): 34193 - 34201.
[Abstract] [Full Text] [PDF]


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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.