TU-tagging: cell type–specific RNA isolation from intact complex tissues
Expressing uracil phosphoribosyltransferase in specific tissues in the fly allows the incorporation of 4-thiouracil into newly synthesized RNA in vivo . The thio-labeled RNA can then be isolated and analyzed by routine procedures allowing the cell type–specific measure of RNA synthesis and decay rat...
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| Veröffentlicht in: | Nature methods 2009-06, Vol.6 (6), p.439-441 |
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| creator | Miller, Michael R Robinson, Kristin J Cleary, Michael D Doe, Chris Q |
| description | Expressing uracil phosphoribosyltransferase in specific tissues in the fly allows the incorporation of 4-thiouracil into newly synthesized RNA
in vivo
. The thio-labeled RNA can then be isolated and analyzed by routine procedures allowing the cell type–specific measure of RNA synthesis and decay rates.
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. |
| doi_str_mv | 10.1038/nmeth.1329 |
| format | Article |
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in vivo
. The thio-labeled RNA can then be isolated and analyzed by routine procedures allowing the cell type–specific measure of RNA synthesis and decay rates.
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.</description><identifier>ISSN: 1548-7091</identifier><identifier>EISSN: 1548-7105</identifier><identifier>DOI: 10.1038/nmeth.1329</identifier><identifier>PMID: 19430475</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>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</subject><ispartof>Nature methods, 2009-06, Vol.6 (6), p.439-441</ispartof><rights>Springer Nature America, Inc. 2009</rights><rights>COPYRIGHT 2009 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Jun 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c565t-b9131ae5e48a5e387f98afd3a5b64437bfcb789f1f248b34808c758baca0d3f13</citedby><cites>FETCH-LOGICAL-c565t-b9131ae5e48a5e387f98afd3a5b64437bfcb789f1f248b34808c758baca0d3f13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nmeth.1329$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nmeth.1329$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19430475$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Miller, Michael R</creatorcontrib><creatorcontrib>Robinson, Kristin J</creatorcontrib><creatorcontrib>Cleary, Michael D</creatorcontrib><creatorcontrib>Doe, Chris Q</creatorcontrib><title>TU-tagging: cell type–specific RNA isolation from intact complex tissues</title><title>Nature methods</title><addtitle>Nat Methods</addtitle><addtitle>Nat Methods</addtitle><description>Expressing uracil phosphoribosyltransferase in specific tissues in the fly allows the incorporation of 4-thiouracil into newly synthesized RNA
in vivo
. The thio-labeled RNA can then be isolated and analyzed by routine procedures allowing the cell type–specific measure of RNA synthesis and decay rates.
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.</description><subject>Animals</subject><subject>Bioinformatics</subject><subject>Biological Microscopy</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>brief-communication</subject><subject>Cellular biology</subject><subject>Drosophila melanogaster</subject><subject>Drosophila melanogaster - chemistry</subject><subject>Drosophila melanogaster - genetics</subject><subject>Gene expression</subject><subject>Genomics</subject><subject>Insects</subject><subject>Life Sciences</subject><subject>Mammals</subject><subject>Nucleic acids</subject><subject>Pentosyltransferases - chemistry</subject><subject>Phosphotransferases</subject><subject>Properties</subject><subject>Proteomics</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA - genetics</subject><subject>RNA - isolation & purification</subject><subject>Scientific method</subject><subject>Separation</subject><subject>Specimen Handling - methods</subject><subject>Staining and Labeling</subject><subject>Thiouracil - analogs & derivatives</subject><subject>Thiouracil - chemistry</subject><subject>Tissue Extracts - chemistry</subject><subject>Tissue Extracts - isolation & 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in vivo
. The thio-labeled RNA can then be isolated and analyzed by routine procedures allowing the cell type–specific measure of RNA synthesis and decay rates.
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.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>19430475</pmid><doi>10.1038/nmeth.1329</doi><tpages>3</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Nature methods, 2009-06, Vol.6 (6), p.439-441 |
| issn | 1548-7091 1548-7105 |
| language | eng |
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| source | MEDLINE; SpringerLink Journals; Nature Journals Online |
| 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 |
| title | TU-tagging: cell type–specific RNA isolation from intact complex tissues |
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