Drosophila Research & Screening Center-Biomedical Technology Research Resource (DRSC-BTRR)

The NIH NIGMS P41-funded DRSC-BTRR helps researchers realize the full potential of Drosophila melanogaster as a model for the study of human health and disease.

View all NIH NIGMS-funded BTRRs and BTDDs

View DRSC-BTRR publications

The DRSC-BTRR develops state-of-the-art tools and methods in three technology areas: (1) Development of technologies for Drosophila cell-based and in vivo studies, (2) Application of technologies for study of mosquito vectors of human diseases, and (3) Development of in vivo proteomics technologies for Drosophila.

We develop technologies through iterative rounds of testing and improvement together with ‘driving biomedical projects’ at collaborating labs that can benefit from the technologies. Current collaborators include experts in cancer therapeutics, rare genetic diseases, and mosquito vectors of infectious diseases.

To further extend the impact of the technologies, we engage in community activities that inform a broad audience and rapidly disseminate technologies. Altogether, we will serve as an integrated, collaborative resource engaging in projects with strong potential for impact in areas that are of interest to several institutes at the US National Institutes of Health.

Point-of-contact for inquiries about DSRC-BTRR technologies and collaborations: Stephanie Mohr

Technology Research & Development (TR&D) focus areas:

TR&D1: Development of CRISPR-based functional genomics technologies for high-throughput screening in Drosophila cultured cells and for use in vivo in Drosophila. Read more about CRISPR knockout in Drosophila cells here.

TR&D2: Development of CRISPR-based functional genomics and other technologies for use in mosquitos, including development of CRISPR screening technologies for use in mosquito cell lines. Read more about mosquito cell technologies here.

TR&D3: Development of proteomics-based technologies for use in vivo in Drosophila, including development of new protein binding and labeling technologies.

Additional components of the DRSC-BTRR include

  • Driving Biomedical Projects (DBPs), which allow us to directly meet the needs of collaborators through iterative technology testing and development
  • Collaboration & Service Projects (CSPs), such as Drosophila cell-based RNAi screens using established technologies
  • Community Engagement, including presentations and workshops aimed at helping the broadest possible research community access DRSC-BTRR technologies
  • Administration & Management, including oversight by NIH NIGMS leadership and scientific advisors to the DRSC-BTRR
  • BioRender illustration of the workflow at the DRSC-BTRR -- from tech development and testing to iterative improvement
  • Illustration by A.L. Ramirez of an Aedes mosquito and a BioRender illustration of the CRISPR cell screening pipeline
  • BioRender illustration evoking the idea of tech development for large-scale screening to find nanobody binders of fly or mosquito proteins
  • BioRender cartoon representing knowhow in the form of notebooks with text and a faint image of a brain

What is the DRSC-BTRR? Plain-language statement of our goals and approaches:

The overall goal of the DRSC-BTRR is to develop new technologies for manipulating genes and proteins in insects or insect cells. Specifically, we are focused on developing technologies that can be applied for research purposes in the fruit fly Drosophila melanogaster, which has long been used to uncover fundamental biological concepts and human disease-relevant information, and in cultured cells from Drosophila or from mosquito species that spread human diseases such as malaria or zika virus disease. To accomplish this--and to maintain a focus on development of technology that truly meets needs--we partner with other laboratories. Among the labs we are currently partnering with are labs focused on the study of rare human diseases, labs focused on understanding the relationship between the microbes that cause diseases and their mosquito hosts, and labs interested to find new treatments for cancer. As part of our efforts, we engage in outreach to research communities that can benefit from our technologies to make sure that they hear about and learn how to use them.

Text illustration that provides a link to the webpage describing our DBPs

Funding: NIGMS P41 GM132087: "Functional genomics resources for the Drosophila and broader research communities" (PI: N. Perrimon | Co-I: S. Mohr)(08/01/2019 - 04/30/2024)

Projects that benefit from our in vivo, cell and/or bioinformatics resources should cite the above grant. Citation is critical to our ability to demonstrate our successful development of resources and outreach to relevant communities so we can continue to receive support.


Recent Publications from the DSRC-BTRR

Raghuvir Viswanatha, Enzo Mameli, Jonathan Rodiger, Pierre Merckaert, Fabiana Feitosa-Suntheimer, Tonya M Colpitts, Stephanie E Mohr, Yanhui Hu, and Norbert Perrimon. 11/24/2021. “Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos.” Nat Commun, 12, 1, Pp. 6825.Abstract
Mosquito-borne diseases present a worldwide public health burden. Current efforts to understand and counteract them have been aided by the use of cultured mosquito cells. Moreover, application in mammalian cells of forward genetic approaches such as CRISPR screens have identified essential genes and genes required for host-pathogen interactions, and in general, aided in functional annotation of genes. An equivalent approach for genetic screening of mosquito cell lines has been lacking. To develop such an approach, we design a new bioinformatic portal for sgRNA library design in several mosquito genomes, engineer mosquito cell lines to express Cas9 and accept sgRNA at scale, and identify optimal promoters for sgRNA expression in several mosquito species. We then optimize a recombination-mediated cassette exchange system to deliver CRISPR sgRNA and perform pooled CRISPR screens in an Anopheles cell line. Altogether, we provide a platform for high-throughput genome-scale screening in cell lines from disease vector species.
Jiunn Song, Arda Mizrak, Chia-Wei Lee, Marcelo Cicconet, Zon Weng Lai, Chieh-Han Lu, Stephanie E. Mohr, Jr Robert V. Farese, and Tobias C. Walther. 9/15/2021. “Identification of two pathways mediating protein targeting from ER to lipid droplets”. Publisher's VersionAbstract
Pathways localizing proteins to their sites of action within a cell are essential for eukaryotic cell organization and function. Although mechanisms of protein targeting to many organelles have been defined, little is known about how proteins, such as key metabolic enzymes, target from the ER to cellular lipid droplets (LDs). Here, we identify two distinct pathways for ER-to-LD (ERTOLD) protein targeting: early ERTOLD, occurring during LD formation, and late ERTOLD, targeting mature LDs after their formation. By using systematic, unbiased approaches, we identified specific membrane-fusion machinery, including regulators, a tether, and SNARE proteins, that are required for late ERTOLD targeting. Components of this fusion machinery localize to LD-ER interfaces and appear to be organized at ER exit sites (ERES) to generate ER-LD membrane bridges. We also identified multiple cargoes for early and late ERTOLD. Collectively, our data provide a new model for how proteins target LDs from the ER.
Yanhui Hu, Sudhir Gopal Tattikota, Yifang Liu, Aram Comjean, Yue Gao, Corey Forman, Grace Kim, Jonathan Rodiger, Irene Papatheodorou, Gilberto Dos Santos, Stephanie E Mohr, and Norbert Perrimon. 2021. “DRscDB: A single-cell RNA-seq resource for data mining and data comparison across species.” Comput Struct Biotechnol J, 19, Pp. 2018-2026.Abstract
With the advent of single-cell RNA sequencing (scRNA-seq) technologies, there has been a spike in studies involving scRNA-seq of several tissues across diverse species including Drosophila. Although a few databases exist for users to query genes of interest within the scRNA-seq studies, search tools that enable users to find orthologous genes and their cell type-specific expression patterns across species are limited. Here, we built a new search database, DRscDB (https://www.flyrnai.org/tools/single_cell/web/), to address this need. DRscDB serves as a comprehensive repository for published scRNA-seq datasets for Drosophila and relevant datasets from human and other model organisms. DRscDB is based on manual curation of Drosophila scRNA-seq studies of various tissue types and their corresponding analogous tissues in vertebrates including zebrafish, mouse, and human. Of note, our search database provides most of the literature-derived marker genes, thus preserving the original analysis of the published scRNA-seq datasets. Finally, DRscDB serves as a web-based user interface that allows users to mine gene expression data from scRNA-seq studies and perform cell cluster enrichment analyses pertaining to various scRNA-seq studies, both within and across species.
Stephanie E Mohr, Sudhir Gopal Tattikota, Jun Xu, Jonathan Zirin, Yanhui Hu, and Norbert Perrimon. 2021. “Methods and tools for spatial mapping of single-cell RNAseq clusters in Drosophila.” Genetics, 217, 4.Abstract
Single-cell RNA sequencing (scRNAseq) experiments provide a powerful means to identify clusters of cells that share common gene expression signatures. A major challenge in scRNAseq studies is to map the clusters to specific anatomical regions along the body and within tissues. Existing data, such as information obtained from large-scale in situ RNA hybridization studies, cell type specific transcriptomics, gene expression reporters, antibody stainings, and fluorescent tagged proteins, can help to map clusters to anatomy. However, in many cases, additional validation is needed to precisely map the spatial location of cells in clusters. Several approaches are available for spatial resolution in Drosophila, including mining of existing datasets, and use of existing or new tools for direct or indirect detection of RNA, or direct detection of proteins. Here, we review available resources and emerging technologies that will facilitate spatial mapping of scRNAseq clusters at high resolution in Drosophila. Importantly, we discuss the need, available approaches, and reagents for multiplexing gene expression detection in situ, as in most cases scRNAseq clusters are defined by the unique coexpression of sets of genes.
Raghuvir Viswanatha, Enzo Mameli, Jonathan Rodiger, Pierre Merckaert, Fabiana Feitosa-Suntheimer, Tonya M. Colpitts, Stephanie E. Mohr, Yanhui Hu, and Norbert Perrimon. 3/30/2021. “Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos [NOTE: A modified final version was published in Nat Comm and is also available here.].” bioRxiv. Publisher's VersionAbstract
Mosquito-borne diseases present a worldwide public health burden. Genome-scale screening tools that could inform our understanding of mosquitos and their control are lacking. Here, we adapt a recombination-mediated cassette exchange system for delivery of CRISPR sgRNA libraries into cell lines from several mosquito species and perform pooled CRISPR screens in an Anopheles cell line. To implement this method, we engineered modified mosquito cell lines, validated promoters and developed bioinformatics tools for multiple mosquito species.Competing Interest StatementThe authors have declared no competing interest.
Xuechun Feng, Víctor López Del Amo, Enzo Mameli, Megan Lee, Alena L Bishop, Norbert Perrimon, and Valentino M Gantz. 2021. “Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes.” Nat Commun, 12, 1, Pp. 2960.Abstract
Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, and avian malaria, posing a constant threat to public health, livestock, companion animals, and endangered birds. While rising insecticide resistance has threatened the control of Culex mosquitoes, advances in CRISPR genome-editing tools have fostered the development of alternative genetic strategies such as gene drive systems to fight disease vectors. However, though gene-drive technology has quickly progressed in other mosquitoes, advances have been lacking in Culex. Here, we develop a Culex-specific Cas9/gRNA expression toolkit and use site-directed homology-based transgenesis to generate and validate a Culex quinquefasciatus Cas9-expressing line. We show that gRNA scaffold variants improve transgenesis efficiency in both Culex quinquefasciatus and Drosophila melanogaster and boost gene-drive performance in the fruit fly. These findings support future technology development to control Culex mosquitoes and provide valuable insight for improving these tools in other species.