Flies as a model for the study of human disease

Rapid forward genetics – isolate mutants throughtransposons or chemical mutagenesis

Rapid construction of transgenic models of human disease

Well established easy systems to drive knockdown/knockout or over expression of gene expression in tissue ortemporal specific patterns

Rapid determination of the molecular basis ofdisease mechanisms

Able to rapidly identify modifier/bypass gene pathways viagenetic screens for enhancersor suppressors of phenotypes

Easy to culture cell lines – very-easy to dsRNA treat genes of interest

GAL4 Tissue specific promoter

X

Transgene UAS OFF

ON

The progeny of this mating will express the transgene in cells also expressing GAL4

GAL4 is a transcriptional activator protein from yeast

The upstream activating sequence is the GAL4 target

GAL4 Tissue specific promoter Transgene UAS

The fly GAL4/UAS binary transgenic expression system

Egg

L1 Larva (juvenile)

L2

L3White (early) pupa

Pupa

Adult

24 hrs

The life cycle of a fly

Drosophila has 4 chromosomes

H. sapiens chromosomes

Embryonic Brain Development

Hartenstein - Atlas of Drosophila Development (1993) Cold Spring Harbour Laboratory Press

Larval/Pupal Brain Development

Hartenstein - Atlas of Drosophila Development (1993) Cold Spring Harbour Laboratory Press

The fly fat body is analogues to adipose tissue, the liver and the haematopoietic system in mammals.

Hotamisilgil (2006) Inflammation and metabolic disorders Nature 444, 860-867

Leopold & Perrimon (2007) Drosophila and the genetics of the internal milieu Nature 450, 186-188

Drosophila oenocytes are analogous to mammalian

hepatocytes

Pandley and Nichols (2011) Human Disease Models in Drosophila melanogaster and the Role of the Fly in Therapeutic Drug Discovery Pharmacological Reviews 63(2)411-436

Testing drug candidates in flies

Throughput

‘Humanized” fly Pex1 mutations

‘Humanized” fly Pex1 mutations

A ‘visible’ screen for peroxisome function in Drosophila eyes

ey – eyeless:GAL4

GMR– Glass Multiple Reporter:GAL4

Pex1*RNAseq

Drosophila Pex1 is expressed throughout development

Pex1*microarray

Drosophila Pex1 is expressed in multiple tissues

Pex3*microarray

Drosophila Pex3 is expressed in multiple tissues

Pex7*microarray

Drosophila Pex7 is expressed highly in the CNS

Drosophila Pex1

Genome wide analysis of Pex1 loss

Genome wide analysis of Pex1 loss

Genome wide analysis of Pex1 loss

Genome wide analysis of Pex1 loss

Loss of Pex1 in flies causes larval lethality

Loss of Pex1 in flies causes a poor growth phenotype

Pex1 mutations do not affect fly musculature

Loss of Pex1 in flies causes a poor growth phenotype

Loss of Pex1 in flies causes severe effects on the Drosophila nervous system

Loss of Pex1 in flies causes

severe effects on the Drosophila nervous system

Loss of Pex1 in flies causes severe effects on the

Drosophila nervous system

High throughput screening

dsRNA library covering 96% of the Drosophila genome

High Throughput screening

Studying Peroxisomes in cultured fly cells

Acknowledgements• U of Alberta

• Simmonds Laboratory• Jing Li• Julie Haskins• Alana Pay

• Rachubinski Laboratory• Jenny Chang• Fred Mast• Robert Tower• Rick Porier

• Dr. Sarah Hughes

• McGill Univeristy• Dr. Nancy Braverman