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Discovery and characterization of novel functional human RNAs by in vitro selection

  • Author(s): Vu, Michael Manh Khang
  • Advisor(s): Luptak, Andrej
  • et al.
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Abstract

The more we learn to read the instructions encoding the functions of all life, the more we realize how much we still have to learn. Our understanding of DNA encoded functions thus far has revolutionized medicine and biological study, but we growing to appreciate how much we still don’t know about the roles of the vast majority of the RNAs that our DNA encodes. My work centers around revealing these undiscovered RNA functions, particularly in humans, and the characterization of these RNAs achieved by harnessing the characteristics of RNA as an evolvable, multi-functional, self-encoded polymer. I used in vitro evolution to isolate and identify RNAs that perform a specified function from the vast landscape of RNA encoded by our genome (genomic SELEX) and again to help define components and properties that grant each of the RNAs that function. I also developed multiplexed methods to chemically probe these RNAs to address the challenge of characterizing the diverse pools of candidates produced by genomic SELEX with current sequencing technology. I discovered the first small molecule aptamers in humans, with now eight known examples of ATP binding RNAs encoded in the human genome. Over the course of their study, I’ve developed CIM-seq and Mini-SELEX and aided in the development of Apta-seq as methods that can be used broadly in the characterization of in vitro selection experiments. Finally, I have found evidence for the existence of a very intriguing ribozyme encoded in our DNA. These contributions inch us closer to understanding the many functions of RNA in vivo, provides topics for further study, and means with which to study them.

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This item is under embargo until October 13, 2020.