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

• Published in: Nature methods Vol. 6; no. 6; pp. 439 - 441
• Main Authors: Doe, Chris Q, Miller, Michael R, Robinson, Kristin J, Cleary, Michael D
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.06.2009; Nature Publishing Group
• Subjects: Animals; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; brief-communication; Cellular biology; Drosophila melanogaster; Drosophila melanogaster - chemistry; Drosophila melanogaster - genetics; Gene expression; Genomics; Insects; Life Sciences; Mammals; Nucleic acids; Pentosyltransferases - chemistry; Phosphotransferases; Properties; Proteomics; Ribonucleic acid; RNA; RNA - genetics; RNA - isolation & purification; Scientific method; Separation; Specimen Handling - methods; Staining and Labeling; Thiouracil - analogs & derivatives; Thiouracil - chemistry; Tissue Extracts - chemistry; Tissue Extracts - isolation & purification; Vertebrates; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.1329

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.