Isolation of mRNA from specific tissues of Drosophila by mRNA tagging

Zhiyong Yang, Howard J. Edenberg, Ronald L. Davis

Research output: Contribution to journalArticle

53 Scopus citations

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

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Isolation of mRNA from specific tissues of Drosophila by mRNA tagging'. Together they form a unique fingerprint.

  • Cite this