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, and is breaking new ground by enabling new studies in mosquito vectors of 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

Interested to use our technologies to help address your biomedical topic of interest? Contact DRSC-BTRR Director Stephanie Mohr

  • 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:

Typical research labs use many technologies to study one or a few biomedical topics. At the DRSC-BTRR, we flip that model. We focus on developing and improving technologies, and we help other labs apply these technologies to study many different topics. What kind of technologies are we working on? We aim to develop new technologies for manipulating genes and proteins in insects or insect cultured 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 either from Drosophila or from mosquitos that spread human diseases such as malaria or zika virus disease. To accomplish this--and to stay focused on technologies that truly meet needs--we partner with laboratories that can benefit from applying the technologies. Among the labs we are currently partnering with are labs focused on the study of rare human genetic diseases, labs interested to find new treatments for cancer, and labs focused on understanding relationships between microbes that cause mosquito-borne diseases and their mosquito hosts. 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. Once technologies are mature, we also publish detailed protocols and provide the materials we have developed, such as DNA plasmids or modified cell lines, to academic and non-profit facilities that specialize in storage and distribution of research materials. Through training, publication of protocols, and transfer of materials to distribution facilities, we make sure that researchers across the US and elsewhere will have easy access to DRSC-BTRR technologies for years to come.

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.

 

Recent Publications from the DSRC-BTRR

Jiunn Song, Arda Mizrak, Chia-Wei Lee, Marcelo Cicconet, Zon Weng Lai, Wei-Chun Tang, Chieh-Han Lu, Stephanie E. Mohr, Robert V. Farese, and Tobias C. Walther. 2022. “Identification of two pathways mediating protein targeting from ER to lipid droplets.” Nature Cell Biol. Publisher's VersionAbstract
Pathways localizing proteins to their sites of action are essential for eukaryotic cell organization and function. Although mechanisms of protein targeting to many organelles have been defined, how proteins, such as metabolic enzymes, target from the endoplasmic reticulum (ER) to cellular lipid droplets (LDs) is poorly understood. Here we identify two distinct pathways for ER-to-LD protein targeting: early targeting at LD formation sites during formation, and late targeting to mature LDs after their formation. Using systematic, unbiased approaches in Drosophila cells, we identified specific membrane-fusion machinery, including regulators, a tether and SNARE proteins, that are required for the late targeting pathway. Components of this fusion machinery localize to LD–ER interfaces and organize at ER exit sites. We identified multiple cargoes for early and late ER-to-LD targeting pathways. Our findings provide a model for how proteins target to LDs from the ER either during LD formation or by protein-catalysed formation of membrane bridges.
Ashley Mae Conard, Nathaniel Goodman, Yanhui Hu, Norbert Perrimon, Ritambhara Singh, Charles Lawrence, and Erica Larschan. 2021. “TIMEOR: a web-based tool to uncover temporal regulatory mechanisms from multi-omics data.” Nucleic Acids Res, 49, W1, Pp. W641-W653.Abstract
Uncovering how transcription factors regulate their targets at DNA, RNA and protein levels over time is critical to define gene regulatory networks (GRNs) and assign mechanisms in normal and diseased states. RNA-seq is a standard method measuring gene regulation using an established set of analysis stages. However, none of the currently available pipeline methods for interpreting ordered genomic data (in time or space) use time-series models to assign cause and effect relationships within GRNs, are adaptive to diverse experimental designs, or enable user interpretation through a web-based platform. Furthermore, methods integrating ordered RNA-seq data with protein-DNA binding data to distinguish direct from indirect interactions are urgently needed. We present TIMEOR (Trajectory Inference and Mechanism Exploration with Omics data in R), the first web-based and adaptive time-series multi-omics pipeline method which infers the relationship between gene regulatory events across time. TIMEOR addresses the critical need for methods to determine causal regulatory mechanism networks by leveraging time-series RNA-seq, motif analysis, protein-DNA binding data, and protein-protein interaction networks. TIMEOR's user-catered approach helps non-coders generate new hypotheses and validate known mechanisms. We used TIMEOR to identify a novel link between insulin stimulation and the circadian rhythm cycle. TIMEOR is available at https://github.com/ashleymaeconard/TIMEOR.git and http://timeor.brown.edu.
Jonathan Zirin, Justin Bosch, Raghuvir Viswanatha, Stephanie E Mohr, and Norbert Perrimon. 2022. “State-of-the-art CRISPR for in vivo and cell-based studies in Drosophila.” Trends Genet, 38, 5, Pp. 437-453.Abstract
For more than 100 years, the fruit fly, Drosophila melanogaster, has served as a powerful model organism for biological and biomedical research due to its many genetic and physiological similarities to humans and the availability of sophisticated technologies used to manipulate its genome and genes. The Drosophila research community quickly adopted CRISPR technologies and, in the 8 years since the first clustered regularly interspaced short palindromic repeats (CRISPR) publications in flies, has explored and innovated methods for mutagenesis, precise genome engineering, and beyond. Moreover, the short lifespan and ease of genetics have made Drosophila an ideal testing ground for in vivo applications and refinements of the rapidly evolving set of CRISPR-associated (CRISPR-Cas) tools. Here, we review innovations in delivery of CRISPR reagents, increased efficiency of cutting and homology-directed repair (HDR), and alternatives to standard Cas9-based approaches. While the focus is primarily on in vivo systems, we also describe the role of Drosophila cultured cells as both an indispensable first step in the process of assessing new CRISPR technologies and a platform for genome-wide CRISPR pooled screens.
Justin A. Bosch and Norbert Perrimon. 2022. “Prime Editing for Precise Genome Engineering in Drosophila.” In Drosophila: Methods and Protocols, edited by Christian Dahmann, Pp. 113 - 134. New York, NY: Springer US. Publisher's VersionAbstract
Editing the Drosophila genome is incredibly useful for gene functional analysis. However, compared to gene knockouts, precise gene editing is difficult to achieve. Prime editing, a recently described CRISPR/Cas9-based technique, has the potential to make precise editing simpler and faster, and produce less errors than traditional methods. Initially described in mammalian cells, prime editing is functional in Drosophila somatic and germ cells. Here, we outline steps to design, generate, and express prime editing components in transgenic flies. Furthermore, we highlight a crossing scheme to produce edited fly stocks in less than 3 months.
Jun Xu, Ah-Ram Kim, Ross W Cheloha, Fabian A Fischer, Joshua Shing Shun Li, Yuan Feng, Emily Stoneburner, Richard Binari, Stephanie E Mohr, Jonathan Zirin, Hidde L Ploegh, and Norbert Perrimon. 2022. “Protein visualization and manipulation in through the use of epitope tags recognized by nanobodies.” Elife, 11.Abstract
Expansion of the available repertoire of reagents for visualization and manipulation of proteins will help understand their function. Short epitope tags linked to proteins of interest and recognized by existing binders such as nanobodies facilitate protein studies by obviating the need to isolate new antibodies directed against them. Nanobodies have several advantages over conventional antibodies, as they can be expressed and used as tools for visualization and manipulation of proteins in vivo. Here, we characterize two short (<15 aa) NanoTag epitopes, 127D01 and VHH05, and their corresponding high-affinity nanobodies. We demonstrate their use in Drosophila for in vivo protein detection and re-localization, direct and indirect immunofluorescence, immunoblotting, and immunoprecipitation. We further show that CRISPR-mediated gene targeting provides a straightforward approach to tagging endogenous proteins with the NanoTags. Single copies of the NanoTags, regardless of their location, suffice for detection. This versatile and validated toolbox of tags and nanobodies will serve as a resource for a wide array of applications, including functional studies in Drosophila and beyond.
Hans M. Dalton, Raghuvir Viswanatha, Ricky Brathwaite Jr., Jae Sophia Zuno, Stephanie E Mohr, Norbert Perrimon, and Clement Y. Chow. 12/4/2021. “A genome-wide CRISPR screen identifies the glycosylation enzyme DPM1 as a modifier of DPAGT1 deficiency and ER stress.” BioRxiv. Publisher's VersionAbstract
Partial loss-of-function mutations in glycosylation pathways underlie a set of rare diseases called Congenital Disorders of Glycosylation (CDGs). In particular, DPAGT1 CDG is caused by mutations in the gene encoding the first step in N-glycosylation, DPAGT1, and this disorder currently lacks effective therapies. To identify potential therapeutic targets for DPAGT1-CDG, we performed CRISPR knockout screens in Drosophila cells for genes associated with better survival and glycoprotein levels under DPAGT1 inhibition. We identified hundreds of candidate genes that may be of therapeutic benefit. Intriguingly, inhibition of the mannosyltransferase Dpm1, or its downstream glycosylation pathways, could rescue two in vivo models of DPAGT1 inhibition and ER stress, even though impairment of these pathways alone usually cause CDGs. While both in vivo models ostensibly cause ER stress (through DPAGT1 inhibition or a misfolded protein), we found a novel difference in fructose metabolism that may indicate glycolysis as a modulator of DPAGT1-CDG. Our results provide new therapeutic targets for DPAGT1-CDG, include the unique finding of Dpm1-related pathways rescuing DPAGT1 inhibition, and reveal a novel interaction between fructose metabolism and ER stress.