Transcript Presentation Slides

Flies as a model for the study of
human disease
Rapid construction of
transgenic models of
human disease
Rapid forward genetics –
isolate mutants through
transposons or chemical
mutagenesis
Rapid determination of the
molecular basis of
disease mechanisms
Easy to culture cell lines –
very-easy to dsRNA treat
genes of interest
Well established easy
systems to drive
knockdown/knockout or
over expression
of gene expression in
tissue or
temporal specific patterns
Able to rapidly identify
modifier/bypass gene
pathways via
genetic screens for
enhancers
or suppressors of
phenotypes
The fly GAL4/UAS binary transgenic
expression system
X
Tissue specific promoter
GAL4
UAS
GAL4 is a transcriptional
activator protein from yeast
Tissue specific promoter
OFF
Transgene
The upstream activating
sequence is the GAL4 target
GAL4
UAS
The progeny of this mating will express the
transgene in cells also expressing GAL4
ON
Transgene
The life cycle of a fly
Adult
Pupa
White (early) pupa
L3
L2
24 hrs
Egg
L1 Larva (juvenile)
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
Drosophila oenocytes are
analogous to mammalian
hepatocytes
Leopold & Perrimon (2007) Drosophila and the genetics of the internal milieu
Nature 450, 186-188
Testing drug candidates in flies
Throughput
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
‘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
Drosophila Pex1 is expressed in
multiple tissues
Pex1
*microarray
Drosophila Pex3 is expressed in
multiple tissues
Pex3
*microarray
Drosophila Pex7 is expressed highly in
the CNS
Pex7
*microarray
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