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TU-tagging: cell type–specific RNA isolation from intact complex tissues

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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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Figure 1: TU-tagging: overview and cell type–specific labeling.
Figure 2: Cell type–specific RNA isolation and analysis.

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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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  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

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  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 to Chris 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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