Combinatorial selection of RNA ligands for complex cellular targets : the RNA liagands-based proteomics.
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 10(14) 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.
Tian
Huicheng
H
Department of Genetics, Center for Genetic and Cellular Therapies, Duke University, Durham, North Carolina 27710, USA. htian@alumnidirector.com
D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D004247:DNA / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D013058:Mass Spectrometry; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020543:Proteome / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D012313:RNA / Q000235:genetics* / Q000378:metabolism*
D008024:Ligands; D009687:Nuclear Proteins; D020543:Proteome; D012313:RNA; D004247:DNA