Drosophila melanogaster Source: Zdenék Berger Mating Egg-laying Embryo larva pupa adult Life Cycle...
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Transcript of Drosophila melanogaster Source: Zdenék Berger Mating Egg-laying Embryo larva pupa adult Life Cycle...
Drosophila natural history• Originated in Africa• Probably spread by human activity• Now found most places where we live• Likes compost, rotting fruit, yeast• Some features conserved, others a reflection of its life
strategy• Harmless (mostly)• Most lab strains derived from isolates collected before 1940’s• Strains collected subsequently have P transposable elements
and can’t easily be used
Model Organisms - a trainspotter’s guide
E. coli Yeast Worm Fly Mouse Human
# Genes 4000 6000 19000 15000 30000? 30000?
Genome (Mb) 4.6 150 3000 300010012
# Neurons 0 (1) 302 105 1011
Where our pet flies live…
Mice - 75c/day150k$/yr
Flies ~ 20k$/yr(consumables and labour)
Can’t be stored frozen :-(Source: John Roote
Fly pushingEarly 1900’s - Drosophila contributes to our understanding of heredity
Mid 1900’s - Grows in popularity among developmental biologists
Homozygous lethal mutations can be kept indefinitely as heterozygous balanced stocks
1970’s - 1980’s - Molecular biology, cloning of Hsp, Hox
1970’s - 1980’s - Large screens for developmental mutants
1982 - Transformation by injection of marked P transposable element into syncytial embryos; transgenic flies identified by marker in F1
1988 - Easy mobilisation of P made possible by stable transposase-producing strains
What’s different?
• More gene redundancy in humans & mammals
• Some organisation of tissues and organs
• Cardiovascular system
• Acquired immunity (antibody response)
• We’re studying them, instead of them studying us
Fly Gene Disruption Projects• Based on transposable element insertion
• Allows further local mutagenesis
• Non-directed - like Venter’s sequencing strategy
• Not random
• ~ 15000 target genes
• include ~ 4000 vital genes
• Requires ~ 1 insertion per 8 kb
• Coverage perhaps 25% of that, more on their way into public domain
Other ways to make “mutants”
• EMS - still has its attractions• Targeted knockouts for reverse genetics• Imprecise excisions for reverse genetics• RNAi for reverse or forward genetics• Deletion kits in defined backgrounds• Ask a fellow flypusher
Getting round early lethality
• GAL4 x UAS-X for targeted expressionCan be used for regulated RNAi expression
Getting round early lethality
• GAL4 x UAS-X for targeted expression• Enhancer/suppressor screens• Mitotic clones (using FLP recombinase)
Getting round early lethality
• GAL4 x UAS-X for targeted expression• Enhancer/suppressor screens• Mitotic clones (using FLP recombinase)• Temperature-sensitive point mutations• RNAi screens in cultured cells
Shared biology - shared diseases
• Cancer
• Ageing
• Neurodegeneration
• Infectious disease
• Models for disease vectors
• Behaviour
• Do flies have disease-gene homologs?
• Do flies have basic cellular processes related to the disease?
• Be nice to a friendly fly geneticist
Flies and “your” disease
The future?
• More insertions• UAS-RNAi collections• SNPs, better mapping of point mutations• Temperature-sensitive alleles for cell biology• Screens take more work in flies than in worms• Some things only possible in flies and not worms -
physiology, some development, some cell biology• “Hopping in” takes about $20k investment, or a
friendly fly lab to drop in on