TU-tagging: cell type-specific RNA isolation from intact complex tissues.
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.
Miller
Michael R
MR
Institute of Neuroscience, Howard Hughes Medical Institute, University of Oregon, Eugene, Oregon, USA.
Robinson
Kristin J
KJ
Cleary
Michael D
MD
Doe
Chris Q
CQ
D000818:Animals; D004331:Drosophila melanogaster / Q000737:chemistry / Q000235:genetics*; D010430:Pentosyltransferases / Q000737:chemistry*; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification*; D013048:Specimen Handling / Q000379:methods; D013194:Staining and Labeling; D013889:Thiouracil / Q000031:analogs & derivatives* / Q000737:chemistry; D014020:Tissue Extracts / Q000737:chemistry* / Q000302:isolation & purification*
D014020:Tissue Extracts; C114719:4-thiouracil; D013889:Thiouracil; D012313:RNA; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase