Isolation of mRNA from specific tissues of Drosophila by mRNA tagging

Zhiyong Yang, Howard Edenberg, Ronald L. Davis

Research output: Contribution to journalArticle

51 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1-9
Number of pages9
JournalNucleic Acids Research
Volume33
Issue number17
DOIs
StatePublished - 2005
Externally publishedYes

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Drosophila
Messenger RNA
Photoreceptor Cells
Poly(A)-Binding Proteins
Genes
Caenorhabditis elegans
Gene Expression Profiling
Microarray Analysis
Drosophila melanogaster
Immunoprecipitation
Antibodies

ASJC Scopus subject areas

  • Genetics

Cite this

Isolation of mRNA from specific tissues of Drosophila by mRNA tagging. / Yang, Zhiyong; Edenberg, Howard; Davis, Ronald L.

In: Nucleic Acids Research, Vol. 33, No. 17, 2005, p. 1-9.

Research output: Contribution to journalArticle

Yang, Zhiyong ; Edenberg, Howard ; Davis, Ronald L. / Isolation of mRNA from specific tissues of Drosophila by mRNA tagging. In: Nucleic Acids Research. 2005 ; Vol. 33, No. 17. pp. 1-9.
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