Drosophila Resources for Modeling Human Diseases

Grant Number: R24OD021997

Research Emphasis/Objectives

Drosophila has been used extensively to model human diseases associated with reduction or loss of gene function. Diseases associated with gene amplification, microduplication, or other changes resulting in over-expression have been more challenging to model. Over-expression models would be extremely valuable, as they would, for example, provide a system for relatively rapid identification of genetic or chemical modifiers of associated phenotypes. Fusion of Cas9 protein to transcriptional activators, in combination with appropriate small guide RNAs (sgRNAs) upstream of a coding sequence, provides a straightforward, effective, and targeted technology for over-expressing genes in Drosophila tissues (CRISPR-OE). This method gives us a unique opportunity to build a much-needed resource to study phenotypes associated with gene over-expression in vivo. The goals of this project are to (1) use our existing Transgenic RNAi Project (TRiP) RNAi and CRISPR sgRNA fly stock production pipeline to develop CRISPR-OE fly models for diseases associated with amplification or duplication; (2) develop CRISPR-OE flies for rate-limiting enzymes, as this information can help elucidate the impact of perturbing metabolic pathways associated with health and disease; and (3) perform an initial characterization of the CRISPR-OE fly stocks, adding to their value for research studies by others. With regard to the first goal, our focus will be on developing fly stock models for genes amplified in cancers or duplicated in syndromes associated with chromosomal microduplications.

TRiP RNAi and TRiP CRISPR reagents result in expected loss-of-function (LOF) and gain-of-function (GOF) phenotypes.

Services Provided

Production of fly stocks with single guide RNAs (sgRNAs) targeting upstream gene regions, such that they are useful for CRISPR over-expression (CRISPR-OE) using the VPR or SAM activation systems. Immediately following production and validation, the fly stocks are shared with the Bloomington Drosophila Stock Center (BDSC) for distribution to the community.

Contact Information

Jonathan Zirin, Ph.D.
Assistant Director of Drosophila RNAi Screening Center (DRSC)/TRiP Functional Genomics Resources
Harvard Medical School
New Research Building, Room 336
77 Avenue Louis Pasteur
Boston, MA 02115
jzirin@genetics.med.harvard.edu

Resource name: Transgenic RNAi Project (TRiP) CRISPR over-expression (TRiP-CRISPR-OE) platform at the DRSC/TRiP Functional Genomics Resources
Fly stock information: fgr.hms.harvard.edu/crispr-fly-stocks-and-vectors
Search and nominate genes for fly stock production: www.flyrnai.org/tools/grna_tracker/web
Fly stock distribution by BDSC: bdsc.indiana.edu/stocks/genome_editing/sgrna.html

Principal Investigator

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

Other/Resource Contacts

Jonathan Zirin, Ph.D.
Assistant Director of DRSC/TRiP Functional Genomics
jzirin@genetics.med.harvard.edu