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Next-generation Drosophila Cell Lines to Elucidate the Cellular Basis of Human Diseases

Grant Number: R24OD019847

Live images of Drosophila cultured cells expressing GFP fusions to proteins localized to specific sub-cellular compartments, generated using a CRISPR/Cas9 knock-in approach.Research Emphasis/Objectives

All diseases result from disturbances at the cellular level, and functional analyses of human diseases increasingly rely on cell biological studies to uncover root causes of disease. Importantly, cell-level analyses are revealing that many diseases affect the function and/or morphology of organelles, signal transduction, homeostasis, or cell growth. For example, cancer is frequently associated with nucleolar hypertrophy and/or altered signaling, neurodegenerative diseases are associated with changes in mitochondria, and many myopathies are associated with lysosomal disorders. These types of defects are readily assayed in cultured cells, given the right cell types, markers, and tools. The availability of resources that enable systematic cell biological analyses in various cell types are critical to assess the functions of genes implicated in diseases. Analysis of the Drosophila genome sequence has revealed striking conservation among fly and human genes. Drosophila orthologs have been identified for about two-thirds of human disease genes, and all major signal transduction pathways are conserved between flies and humans. The full potential of Drosophila cell lines, particularly for disease-related studies, has not yet been realized, as the repertoire of cell lines is limited, as is the availability of live-cell reporters and mutant cell lines. To overcome these unmet needs, we are using exciting new advances to generate reagents that will enable the community to fully exploit the power of Drosophila cell lines to address disease-related questions. The goals of this work are to (1) generate new cell lines derived from specific Drosophila cell types, using a technique pioneered by Dr. Simcox; (2) use CRISPR-Cas9 genome engineering to develop a resource of cell lines expressing fluorescent markers that "light up" specific subcellular components, such as the nucleus or mitochondria; and (3) use CRISPR-Cas9 gene editing to generate "knockout" cell lines for cell-based screens. For this last aim, we are focusing on knockout of fly orthologs of tumor suppressor genes to facilitate synthetic lethal screens, which have the potential to identify new targets for cancer treatment.

Services Provided

We build CRISPR-modified Drosophila cell lines and derive new Drosophila cell lines using a tissue-specific RasV12 approach. These cell line resources are shared with the Drosophila Genomics Resource Center (DGRC) for distribution to the community.

Contact Information

Stephanie Mohr, Ph.D.
Director of Drosophila RNAi Screening Center/TRiP Functional Genomics Resources
Harvard Medical School
New Research Building, Room 336
77 Avenue Louis Pasteur
Boston, MA 02115
stephanie_mohr@hms.harvard.edu

Resource name: Drosophila RNAi Screening Center (DRSC) modified cell line collection at DRSC/TRiP Functional Genomics Resources
Website (CRISPR-modified cell line information at the DRSC): fgr.hms.harvard.edu/crispr-modified-cell-lines
Website (new and CRISPR-modified cell line distribution by DGRC): dgrc.bio.indiana.edu/cells/Catalog

Co-Principal Investigators

Norbert Perrimon, Ph.D.
Harvard Medical School
Phone: 617-432-7672
perrimon@genetics.med.harvard.edu

Amanda Simcox, Ph.D.
Ohio State University
Phone: 614-292-8857
simcox.1@osu.edu

Other/Resource Contacts

Stephanie Mohr, Ph.D.
Co-Investigator and Director of DRSC/TRiP Functional Genomics
stephanie_mohr@hms.harvard.edu