1. Genetic annotation of gain-of-function screens using RNA interference and in situ hybridization of candidate genes in the Drosophila wing.
- Author
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Molnar C, Casado M, López-Varea A, Cruz C, and de Celis JF
- Subjects
- Animals, Gene Expression Regulation, Developmental, Genetic Association Studies, In Situ Hybridization, Mutagenesis, Insertional genetics, Mutation, Phenotype, RNA Interference, Receptors, Notch genetics, Receptors, Notch metabolism, DNA Transposable Elements genetics, Drosophila Proteins genetics, Drosophila melanogaster genetics, Drosophila melanogaster growth & development, Drosophila melanogaster metabolism, Transcription Factors genetics, Wings, Animal cytology, Wings, Animal metabolism
- Abstract
Gain-of-function screens in Drosophila are an effective method with which to identify genes that affect the development of particular structures or cell types. It has been found that a fraction of 2-10% of the genes tested, depending on the particularities of the screen, results in a discernible phenotype when overexpressed. However, it is not clear to what extent a gain-of-function phenotype generated by overexpression is informative about the normal function of the gene. Thus, very few reports attempt to correlate the loss- and overexpression phenotype for collections of genes identified in gain-of-function screens. In this work we use RNA interference and in situ hybridization to annotate a collection of 123 P-GS insertions that in combination with different Gal4 drivers affect the size and/or patterning of the wing. We identify the gene causing the overexpression phenotype by expressing, in a background of overexpression, RNA interference for the genes affected by each P-GS insertion. Then, we compare the loss and gain-of-function phenotypes obtained for each gene and relate them to its expression pattern in the wing disc. We find that 52% of genes identified by their overexpression phenotype are required during normal development. However, only in 9% of the cases analyzed was there some complementarity between the gain- and loss-of-function phenotype, suggesting that, in general, the overexpression phenotypes would not be indicative of the normal requirements of the gene.
- Published
- 2012
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