Skip to main content
SpringerLink
Log in

In situ detection of miRNAs in animal embryos using LNA-modified oligonucleotide probes

  • Brief Communication
  • Published:

From Nature Methods

View current issue Submit your manuscript

Abstract

MicroRNAs (miRNAs) are 20–23 nucleotide (nt) RNA molecules that regulate gene expression post-transcriptionally. A key step toward understanding the function of the hundreds of miRNAs identified in animals is to determine their expression during development. Here we performed a detailed analysis of conditions for in situ detection of miRNAs in the zebrafish embryo using locked nucleic acid (LNA)-modified DNA probes and report expression patterns for 15 miRNAs in the mouse embryo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Figure 1: miRNA detection in zebrafish embryos using LNA-modified DNA probes.
Figure 2: Expression of 15 miRNAs in 9.5- and 10.5-d.p.c. mouse embryos.

Similar content being viewed by others

References

  1. Griffiths-Jones, S. Nucleic Acids Res. 32, D109–D111 (2004).

    Article  CAS  Google Scholar 

  2. Bentwich, I. et al. Nat. Genet. 37, 766–770 (2005).

    Article  CAS  Google Scholar 

  3. Berezikov, E. et al. Cell 120, 21–24 (2005).

    Article  CAS  Google Scholar 

  4. Wienholds, E. & Plasterk, R.H. FEBS Lett. 579, 5911–5922 (2005).

    Article  CAS  Google Scholar 

  5. Giraldez, A.J. et al. Science 308, 833–838 (2005).

    Article  CAS  Google Scholar 

  6. Zhao, Y., Samal, E. & Srivastava, D. Nature 436, 214–220 (2005).

    Article  CAS  Google Scholar 

  7. Lu, J. et al. Nature 435, 834–838 (2005).

    Article  CAS  Google Scholar 

  8. Wienholds, E. et al. Science 309, 310–311 (2005).

    Article  CAS  Google Scholar 

  9. Vester, B. & Wengel, J. Biochemistry 43, 13233–13241 (2004).

    Article  CAS  Google Scholar 

  10. Mansfield, J.H. et al. Nat. Genet. 36, 1079–1083 (2004).

    Article  CAS  Google Scholar 

  11. Sempere, L.F. et al. Genome Biol. 5, R13 (2004).

    Article  Google Scholar 

  12. Thomson, J.M., Parker, J., Perou, C.M. & Hammond, S.M. Nat. Methods 1, 47–53 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Exiqon for kindly providing the miRCURY probes. We thank B. Ason, M. Tijsterman and E. Cuppen for reading the manuscript critically. This work was supported by the Council for Earth and Life Sciences of the Netherlands Organization for Scientific Research.

Author information

Authors and Affiliations

  1. Hubrecht Laboratory, Center for Biomedical Genetics, Uppsalalaan 8, Utrecht, 3584 CT, The Netherlands

    Wigard P Kloosterman, Erno Wienholds, Ewart de Bruijn & Ronald H A Plasterk

  2. Wilhelm Johannsen Centre for Functional Genome Research, Institute of Medical Biochemistry and Genetics, University of Copenhagen, Blegdamsvej 3, DK-2200, Denmark

    Sakari Kauppinen

Authors
  1. Wigard P Kloosterman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Erno Wienholds
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ewart de Bruijn
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sakari Kauppinen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ronald H A Plasterk
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ronald H A Plasterk.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Detection of miR-124a, miR-122a and miR-206 with DIG-labeled DNA and LNA probes in 72h zebrafish embryos. (PDF 602 kb)

Supplementary Fig. 2

Determination of the optimal hybridization temperature for in situ detection of miRNAs in the zebrafish embryo. (PDF 3240 kb)

Supplementary Fig. 3

Determination of the optimal hybridization time for in situ detection of miRNAs in the zebrafish embryo. (PDF 1215 kb)

Supplementary Fig. 4

Assessment of the specificity of LNA-modified probes using perfectly matched and mismatched 14-mer and 22-mer LNA-modified probes for miR-124a and miR-206. (PDF 1060 kb)

Supplementary Fig. 5

Detection of miR-183 and miR-217 using LNA-modified probes designed to target the mature, star and loop sequences of the miRNA precursor. (PDF 1303 kb)

Supplementary Fig. 6

In situ hybridizations of miRNAs in Xenopus tropicalis embryos. (PDF 493 kb)

Supplementary Table 1

Short probes, mismatch probes and star and loop probes used in this study. (PDF 79 kb)

Supplementary Table 2

In situ probes and detection of miRNAs in mouse. (PDF 76 kb)

Supplementary Methods (PDF 76 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kloosterman, W., Wienholds, E., de Bruijn, E. et al. In situ detection of miRNAs in animal embryos using LNA-modified oligonucleotide probes. Nat Methods 3, 27–29 (2006). https://doi.org/10.1038/nmeth843

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth843

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation