Abstract
It was only relatively recently discovered that nucleic acids participate in a variety of biological functions, besides the storage and transmission of genetic information. Quite apart from the nucleotide sequence, it is now clear that the structure of a nucleic acid plays an essential role in its functionality, enabling catalysis and specific binding reactions. In vitro selection and evolution strategies have been extremely useful in the analysis of functional RNA and DNA molecules, helping to expand our knowledge of their functional repertoire and to identify and optimize DNA and RNA molecules with potential therapeutic and diagnostic applications. The great progress made in this field has prompted the development of ex vivo methods for selecting functional nucleic acids in the cellular environment. This review summarizes the most important and most recent applications of in vitro and ex vivo selection strategies aimed at exploring the therapeutic potential of nucleic acids.
Highlights
Nucleic acids, RNA, are extremely versatile molecules
It was in the 1960s when Spiegelman’s group first observed evolution in vitro [1]. These authors reported that changes in the RNA genome of the Qβ bacteriophage during replication led to the formation of RNA molecules that were more efficiently copied by the viral replicase
Primer binding sites (PBS), flanking the variable region are incorporated during the design of the starting population to facilitate the amplification of desired molecules, this limits the structural diversity of the RNA populations to specific conformations [14]
Summary
RNA, are extremely versatile molecules. Apart from their role as carriers of genetic information they can express a phenotype, e.g., they may show a catalytic activity, have a specific binding function, or have the capacity to recruit specific molecules. Over the last 20 years, advances in molecular biology and biotechnology have seen the development of methods that allow the effect of such naturally arising variation to be mimicked in the laboratory It was in the 1960s when Spiegelman’s group first observed evolution in vitro [1]. These authors reported that changes in the RNA genome of the Qβ bacteriophage during replication led to the formation of RNA molecules that were more efficiently copied by the viral replicase. New selection methods – known as ex vivo selection procedures – are being used to identify molecules that target viruses, subcellular fractions and even whole cells These techniques overcome some of the limits imposed by in vitro technology and provide new environments and conditions in which to explore the properties and functions of nucleic acids. This review highlights the most recent advances in in vitro and ex vivo selection procedures for nucleic acids, and discusses their potential application in biomedicine
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