Profiling patterned transcripts in Drosophila embryos

Here we describe a high-throughput screen to isolate transcripts with spatially restricted patterns of expression in early embryos. Our approach utilizes robotic automation for rapid analysis of sequence-selected cDNAs in a whole-mount in situ hybridization assay. We determined the spatial distribut...

Full description

Saved in:
Bibliographic Details
Published in:Genome research 2002-07, Vol.12 (7), p.1040-1047
Main Authors: Simin, Karl, Scuderi, Anne, Reamey, James, Dunn, Diane, Weiss, Robert, Metherall, James E, Letsou, Anthea
Format: Article
Language:English
Subjects:
Animals
DNA, Complementary - classification
DNA, Complementary - genetics
DNA, Complementary - physiology
Drosophila - embryology
Drosophila - genetics
Embryo, Nonmammalian - chemistry
Gene Expression Profiling - methods
Genes, Insect - genetics
Letter
RNA, Messenger - genetics
Sequence Analysis, DNA - methods
Transcription, Genetic
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Here we describe a high-throughput screen to isolate transcripts with spatially restricted patterns of expression in early embryos. Our approach utilizes robotic automation for rapid analysis of sequence-selected cDNAs in a whole-mount in situ hybridization assay. We determined the spatial distribution of a random collection of 778 different genes from an embryonic cDNA library and show that a significant fraction of these exhibit patterned profiles of expression. In addition, gene ontology studies revealed groups of gene products exhibiting shared expression patterns, providing new insights into the largely overlooked effector molecules that function in development. As described in this paper, automated hybridization to whole-mount embryos in situ proved to be straightforward and provided us with a very powerful method for the global survey of gene expression in early embryos. From the perspective of biological significance, our finding that many spatially restricted transcripts correspond to loci encoding novel transcripts that have not been previously identified in nearly saturating genetic screens for maternal effect and zygotic lethals is particularly notable.
ISSN:1088-9051
1054-9803
1549-5469
DOI:10.1101/gr.84402