title: Isolation of mRNA from specific tissues of Drosophila by mRNA tagging.
abstract: To study the function of specific cells or tissues using genomic tools like microarray analyses, it is highly desirable to obtain mRNA from a homogeneous source. However, this is particularly challenging for small organisms, like Caenorhabditis elegans and Drosophila melanogaster. We have optimized and applied a new technique, mRNA tagging, to isolate mRNA from specific tissues of D.melanogaster. A FLAG-tagged poly(A)-binding protein (PABP) is expressed in a specific tissue and mRNA from that tissue is thus tagged by the recombinant PABP and separated from mRNA in other tissues by co-immunoprecipitation with a FLAG-tag specific antibody. The fractionated mRNA is then amplified and used as probe in microarray experiments. As a test system, we employed the procedures to identify genes expressed in Drosophila photoreceptor cells. We found that most known photoreceptor cell-specific mRNAs were identified by mRNA tagging. Furthermore, at least 11 novel genes have been identified as enriched in photoreceptor cells. mRNA tagging is a powerful general method for profiling gene expression in specific tissues and for identifying tissue-specific genes.
authors.affiliation: Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Yang
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Edenberg
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Davis
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D020869:Gene Expression Profiling / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009842:Oligopeptides; D010455:Peptides / Q000235:genetics; D017956:Photoreceptor Cells, Invertebrate / Q000378:metabolism; D039102:Poly(A)-Binding Proteins / Q000235:genetics; D015347:RNA Probes / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D005404:Fixatives; D009842:Oligopeptides; D010455:Peptides; D039102:Poly(A)-Binding Proteins; D015347:RNA Probes; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D005557:Formaldehyde; C085706:FLAG peptide