Nucleic acid-based fluorescent probes and their analytical potential |
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Authors: | Juskowiak Bernard |
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Institution: | (1) Faculty of Chemistry, A. Mickiewicz University, Grunwaldzka 6, 60-780 Poznan, Poland |
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Abstract: | It is well known that nucleic acids play an essential role in living organisms because they store and transmit genetic information
and use that information to direct the synthesis of proteins. However, less is known about the ability of nucleic acids to
bind specific ligands and the application of oligonucleotides as molecular probes or biosensors. Oligonucleotide probes are
single-stranded nucleic acid fragments that can be tailored to have high specificity and affinity for different targets including
nucleic acids, proteins, small molecules, and ions. One can divide oligonucleotide-based probes into two main categories:
hybridization probes that are based on the formation of complementary base-pairs, and aptamer probes that exploit selective
recognition of nonnucleic acid analytes and may be compared with immunosensors. Design and construction of hybridization and
aptamer probes are similar. Typically, oligonucleotide (DNA, RNA) with predefined base sequence and length is modified by
covalent attachment of reporter groups (one or more fluorophores in fluorescence-based probes). The fluorescent labels act
as transducers that transform biorecognition (hybridization, ligand binding) into a fluorescence signal. Fluorescent labels
have several advantages, for example high sensitivity and multiple transduction approaches (fluorescence quenching or enhancement,
fluorescence anisotropy, fluorescence lifetime, fluorescence resonance energy transfer (FRET), and excimer-monomer light switching).
These multiple signaling options combined with the design flexibility of the recognition element (DNA, RNA, PNA, LNA) and
various labeling strategies contribute to development of numerous selective and sensitive bioassays. This review covers fundamentals
of the design and engineering of oligonucleotide probes, describes typical construction approaches, and discusses examples
of probes used both in hybridization studies and in aptamer-based assays. |
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