TU-tagging: cell type–specific RNA isolation from intact complex tissues (original) (raw)

Nature Methods volume 6, pages 439–441 (2009)Cite this article

Abstract

We found that the combination of spatially restricted uracil phosphoribosyltransferase (UPRT) expression with 4-thiouracil delivery can be used to label and purify cell type–specific RNA from intact complex tissues in Drosophila melanogaster. This method is useful for isolating RNA from cell types that are difficult to isolate by dissection or dissociation methods and should work in many organisms, including mammals and other vertebrates.

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References

  1. Tomancak, P. et al. Genome Biol. 8, R145 (2007).
    Article Google Scholar
  2. Wang, Z., Gerstein, M. & Snyder, M. Nat. Rev. Genet. 10, 57–63 (2009).
    Article CAS Google Scholar
  3. Schadt, E.E., Monks, S.A. & Friend, S.H. Biochem. Soc. Trans. 31, 437–443 (2003).
    Article CAS Google Scholar
  4. Roy, P.J. et al. Nature 418, 975–979 (2002).
    Article CAS Google Scholar
  5. Tanke, H.J. & van der Keur, M. Trends. Biotechnol. 11, 55–62 (1993).
    Article CAS Google Scholar
  6. Doyle, J.P. et al. Cell 135, 749–762 (2008).
    Article CAS Google Scholar
  7. Heiman, M. et al. Cel 135, 738–748 (2008).
    Article CAS Google Scholar
  8. Nelson, S.B., Hempel, C. & Sugino, K. Curr. Opin. Neurobiol. 16, 571–576 (2006).
    Article CAS Google Scholar
  9. Cleary, M.D. et al. Nat. Biotechnol. 23, 232–237 (2005).
    Article CAS Google Scholar
  10. Dolken, L. et al. RNA 14, 1959–1972 (2008).
    Article Google Scholar
  11. Bainton, R.J. et al. Cell 123, 145–156 (2005).
    Article CAS Google Scholar
  12. Ebens, A.J. et al. Cell 74, 15–27 (1993).
    Article CAS Google Scholar
  13. Awasaki, T. et al. Neuron 50, 855–867 (2006).
    Article CAS Google Scholar
  14. Rinn, J.L. et al. Genes Dev. 22, 303–307 (2008).
    Article CAS Google Scholar
  15. Zeiner, G.M. et al. Methods Mol. Biol. 419, 135–146 (2008).
    Article CAS Google Scholar

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Acknowledgements

We thank C. Canestro, T. Herman, Z. Lewis and S. O'Rourke for comments on the manuscript, and C. Cabernard for help with Imaris. M.R.M. was supported by a US National Science Foundation predoctoral fellowship, M.D.C. was supported by a National Institutes of Health National Research Service Award postdoctoral fellowship, and C.Q.D. was supported by National Institutes of Health HD27056 and the Howard Hughes Medical Institute.

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Author notes

  1. Michael D Cleary
    Present address: Present address: School of Natural Sciences, University of California, Merced, California, USA.,

Authors and Affiliations

  1. Institute of Neuroscience, Institute of Molecular Biology, Howard Hughes Medical Institute, University of Oregon, Eugene, Oregon, USA
    Michael R Miller, Kristin J Robinson, Michael D Cleary & Chris Q Doe

Authors

  1. Michael R Miller
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  2. Kristin J Robinson
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  3. Michael D Cleary
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  4. Chris Q Doe
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Correspondence toChris Q Doe.

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Miller, M., Robinson, K., Cleary, M. et al. TU-tagging: cell type–specific RNA isolation from intact complex tissues.Nat Methods 6, 439–441 (2009). https://doi.org/10.1038/nmeth.1329

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