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The Transgenic Study of Mus musculus

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create mouse models of human diseases. Two Transgenic Techniques ... a transgenic mouse ... of DNA on chromosome 17 and mice developed short tails ... – PowerPoint PPT presentation

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Title: The Transgenic Study of Mus musculus


1
The Transgenic Study of Mus musculus
  • by Brian Stephenson
  • and Arthur Musarra

2
After Implantation.
  • Placenta- nutrients through blood supply and
    waste removal
  • Gastrulation cells differentiate to germ layers
  • Endoderm
  • Ectoderm
  • Mesoderm
  • 21 days later birth
  • 5-6 weeks following ready to breed

3
Uses of Transgenic Tools
  • determine the function of gene products
  • characterize genetic regulatory regions
  • est. links between mutant phenotypes and
    particular transcriptional units to identify a
    cloned gene
  • create mouse models of human diseases

4
Two Transgenic Techniques
  • Nuclear Injection
  • Targeted Mutagenesis

5
Nuclear Injection
  • Injection of foreign DNA into a pronucleus which
    is placed back into a female oviduct for
    development
  • This process creates a transgenic mouse
  • Transgene comes from animals, plant, microbial,
    DNA synthesis

6
Limitations to Nuclear Injection
  • Only add not subtract genetic material
  • Cannot target insertion of DNA to specific
    genomic locations

7
Targeted Mutagenesis
  • Replaces current allele with a new allele of
    different function
  • undifferentiated
  • altered cells, known
  • as embryonic stem
  • cells, injected into the
  • blastocoele
  • This is known
  • as transfection

8
Targeted Mutagenesis
  • Specific allele may be replaced with new allele
  • These alleles may be knockout constructs

9
Determining Gene Function
  • Using transgenic technology geneticists showed
    that cloned SRY gene encodes male sex organs
    without a Y chromosome

10
Models of Disease
  • non-cancerous animal cells have a very low level
    of myc gene expression
  • gene discovered in the myelocytomatosis chicken
    retrovirus
  • exposure generated tumors at high levels of gene
    expression

11
Models of Disease
  • To test myc gene used transgenic line of
  • immunoglobulin- producing B-Cells
  • tissue specific promoters - mammilary gland
  • general tissue promoters
  • Conclusion myc gene alone does not cause cells
    to become cancerous but can work with other
    mutations to cause cancerous growth

12
Models of Disease
  • use targeted mutagenesis to knockout genes for
    disease models
  • ex CFTR gene for human cystic fibrosis
  • enables further research to be done to correct
    loss of gene functioning in animal models

13
To Study Genetic Regulatory Regions
  • clone DNA with gene of interest and reg. region
  • exchange DNA coding sequence for reporter lacZ
    gene from E. coli
  • make several different forms of transgene for
    possible regulatory regions
  • Tract and compare natural gene to map regulatory
    function

14
Linking Gene to Phenotype
  • a study used X-ray induced mutations that deleted
    a portion of DNA on chromosome 17 and mice
    developed short tails
  • linked pme75 gene to tail length
  • added pme75 to transgene construct and introduced
    to short tail mutants, offspring developed normal
    tails
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