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Related Experiment Videos

Targeting Functional Noncoding RNAs.

Madzia P Crossley1,2, Torsten Krude3

  • 1Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK.

Methods in Molecular Biology (Clifton, N.J.)
|April 2, 2017
PubMed
Summary

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Morpholino antisense oligonucleotides effectively inactivate small noncoding RNAs crucial for DNA replication in both vertebrate Y RNAs and nematode stem-bulge RNAs. This study details methods for targeting these essential noncoding RNAs.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Noncoding RNAs play vital roles in cellular metabolism, including DNA replication.
  • Targeting noncoding RNAs is essential for understanding their functions.

Purpose of the Study:

  • To discuss challenges and present protocols for inactivating functional noncoding RNAs using Morpholino antisense oligonucleotides.
  • To demonstrate the efficacy of this method for small noncoding RNAs involved in DNA replication.

Main Methods:

  • Design, preparation, and efficacy controls of Morpholino antisense oligonucleotides.
  • Delivery of Morpholino antisense oligonucleotides into vertebrate and nematode embryos.
  • Functional inactivation of Y RNAs (vertebrates) and stem-bulge RNAs (nematodes).
Keywords:
Caenorhabditis elegansDanio rerioDominant-negative inhibitionFunctional assay to test Morpholino efficacy and specificityFunctional inhibitionMorpholino deliveryMorpholino designNoncoding RNA domainsNoncoding RNAsXenopus laevis

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Main Results:

  • Successful functional inactivation of small noncoding RNAs essential for DNA replication.
  • Demonstrated efficacy of Morpholino antisense oligonucleotides in both vertebrate and nematode models.
  • Established protocols for targeting and delivering these oligonucleotides.

Conclusions:

  • Morpholino antisense oligonucleotides are a viable tool for functional inactivation of noncoding RNAs.
  • This approach is effective for studying noncoding RNAs involved in DNA replication.
  • Protocols provided facilitate research in noncoding RNA biology and developmental processes.