The Drosophila RNAi Screening Center is principally funded by a grant from the National Institute of General Medical Sciences, a division of the National Institutes of Health. The project was launched with seed funds from Howard Hughes Medical Institute and Harvard Medical School. The DRSC is also participating in the collaborative Dana Farber/ Harvard Cancer Center (DF/HCC) RNAi Core. The DF/HCC is an NCI designated Comprehensive Cancer Center.

Current Grants

NIGMS R01 GM067761:
"Functional Genomic analysis using RNAi screen in Drosophila"
Funding Period: 5/01/03 - 11/30/15

Grant Summary: Funding provided in the first cycle of this grant helped to establish at Harvard Medical School the Drosophila RNAi Screening Center (DRSC). The purpose of the facility was to provide the community with a unique infrastructure and expertise for high-throughput RNAi screens (RNAi HTS) in Drosophila cell lines using a genome-wide collection of double stranded RNAs (dsRNAs). The DRSC has been a resounding success. In less than 3 years, 75 applications from 43 different institutions have been approved by the center, 47 screens have been completed and 11 primary papers detailing the screens have been already published. The vast experience gained during these first years has been instrumental in shaping our view on how to build upon our initial goals and expand the scope and technology of RNAi HTS in Drosophila. To face those new challenges and develop the DRSC into an integrated center for technology transfer in functional genomics, we propose the following: First, the off-target effects observed with long dsRNAs will be addressed through a re-design of our library and the ability to offer two or more dsRNAs to confirm the specificity of RNAi phenotypes. Second, we will implement a number of key improvements to expand the range of functional screens available at the DRSC. We will offer sub-collections of dsRNAs against a variety of large gene families such as transcription factors or cytoskeletal proteins for more focused screens, and add miRNA and cDNA constructs over-expression libraries for gain of function studies in Drosophila cell- based assays. Third, to meet the growing demand on performing more sophisticated screens with greater biological relevance, we have added the ability to perform RNAi HTS in primary Drosophila cell cultures and have acquired an ultra high-throughput confocal, imaging platform that will be ideal for high content imaging screens. Fourth, we address the need of screeners to handle vast amount of data and the challenge to analyze them by planning a major upgrade of our database and developing powerful computational approaches for data analysis and data mining. Lastly, we provide two practical solutions to validate the results of the primary RNAi screen: the ability to perform focused RNAi screens in mammalian cells with siRNAs targeting human orthologs of genes identified in the Drosophila screen, and the generation of custom short hairpin RNA transgenic flies for rapid in vivo validation.

NIGMS R01 GM084947:
"Drosophila Transgenic RNAi Resource Project"

Grant Summary: In Drosophila, arguably the best-understood multicellular organism and a proven model system for human diseases, less than half of the 15,185 annotated genes have mutations, and many fewer have readily detectable phenotypes. Our lack of functional information on the majority of the genes, also referred to as the "Phenotype Gap", does not indicate that these genes have no function but rather that, as experimentalists, we have been unable to either assay their roles or resolve the issue of functional redundancy. Conditional expression of hairpin constructs in Drosophila has emerged in recent years as the method of choice to fill in the "Phenotype Gap", as well as to overcome the issues associated with gene pleiotropy. Using transgenic RNAi it is now possible to disrupt the activity of single genes with a spatial and temporal resolution that is impossible or exceedingly difficult using classical genetic methods. Here, we propose to build a resource of 6,250 transgenic RNAi lines, the "Transgenic RNAi Resource Project", using improved methodology developed in our laboratory. The lines will be established and validated at the Drosophila RNAi Screening Center (DRSC) and transferred to the Bloomington Drosophila Stock Center (BDSC) to be made freely available to the community. We anticipate this resource to be built, tested and transferred in its entirety to the BDSC over a four-year period. This collection of transgenic RNAi lines will be invaluable to address a myriad of questions in biology and medicine, including, but not limited to, cell biology, signal transduction and cancer, the etiology of congenital malformations, neurodegenerative studies, and behavior.

R24 RR032668:
"Development of validated Drosophila in vivo RNAi models of human diseases"

Grant Summary: Remarkable conservation exists between Drosophila and humans at the level of genes and gene functions. For example, there are identifiable fly orthologs for about two-thirds of human disease genes and all of the major signal transduction pathways have been conserved between flies and humans. The availability of RNAi for gene-specific knockdown of mRNA levels has accelerated the pace at which we can undertake the type of molecular genetic analyses that make Drosophila such a powerful system. However, despite the impressive set of tools currently available, locating and/or generating Drosophila disease models can be time consuming, and in most cases, well-characterized loss-of-function alleles or validated RNAi strains are not available. The proposed project would build upon our existing expertise and infrastructure to produce a large collection of in vivo Drosophila RNAi models of human disease, the HuDis-TRiP Resource of Disease Models. This resource will be unique in its large scale, targeting approximately 900 Drosophila orthologs of human disease genes, and in the high level of quality control applied to each RNAi fly stock. For human disease genes for which loss-of-function of the gene is associated with a disease state, HuDis-TRiP fly stocks are likely to serve as disease models by mimicking the human disease state at the cell, tissue and/or organism level. Specifically, we propose to: Aim 1, compile a prioritized list of human disease genes using the Online Mendelian Inheritance in Man database and a community nomination process; Aim 2, identify 900 high-confidence fly orthologs of these genes; make two transgenic Drosophila RNAi fly stocks per gene; perform phenotypic characterization, quantitative PCR and rescue to validate the resource; and Aim 3, further characterize the most promising models. Notably, our initial list of conserved disease-associated genes includes genes relevant to nearly all NIH Institutes. In keeping with our commitment to community-based efforts, all RNAi fly stocks will be rapidly transferred to a public stock center and all datasets will be made available through our own database, website (www.flyrnai.org) and other databases. We anticipate that this resource will be widely used by the community, as it will allow scientists to immediately begin their studies with high quality, validated disease models in a powerful genetic system.