Abstract

Oligonucleotide microarrays are widely used in various biological studies. In this review, application of oligonucleotide microarrays for identifying binding sites and probing structure of RNAs is described. Deep sequencing allows fast determination of DNA and RNA sequence. High-throughput methods for determination of secondary structures of RNAs have also been developed. Those methods, however, do not reveal binding sites for oligonucleotides. In contrast, microarrays directly determine binding sites while also providing structural insights. Microarray mapping can be used over a wide range of experimental conditions, including temperature, pH, various cations at different concentrations and the presence of other molecules. Moreover, it is possible to make universal microarrays suitable for investigations of many different RNAs, and readout of results is rapid. Thus, microarrays are used to provide insight into oligonucleotide sequences potentially able to interfere with biological function. Better understanding of structure–function relationships of RNA can be facilitated by using microarrays to find RNA regions capable to bind oligonucleotides. That information is extremely important to design optimal sequences for antisense oligonucleotides and siRNA because both bind to single-stranded regions of target RNAs.

Highlights

  • Microarrays contain a library of oligonucleotides or polynucleotides with each spotted in a defined location

  • Often probes bound to specific regions in all three mRNAs, but strong hybridization was not correlated with probe base composition, presumably because of secondary structure in the target RNA

  • DNA oligonucleotides identified as strong binders in microarray experiments inhibited translation of their cognate targets in Xenopus egg extract and strongly reduced synthesis of the targeted cyclins, whereas those oligonucleotides with weak binding to microarrays produced no or little inhibition

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Summary

Introduction

Microarrays contain a library of oligonucleotides or polynucleotides with each spotted in a defined location. Based on the studies of interactions of model oligonucleotides, the Southern group demonstrated that (i) probes bind to single-stranded regions of RNA and the presence of mismatches between probe and RNA sequence reduces binding intensity, (ii) strong hybridization Can be observed when only a part of long probes canonically base-pairs to target RNA and (iii) the presence of secondary structural motifs adjacent to single-stranded regions influences hybridization.

Results
Conclusion

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