Lecture 7b - BIO 311 RNAi in C elegans Model Organisms in...

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BIO 311 RNAi in C. elegans
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Model Organisms in Biomedical Research Rats, Mice and other Rodents ≤ 97% Cats and Dogs ≤ 1% Fish, Frogs, Reptiles, Birds Non-human Primates ≤ 0.3% (GB, 2004)
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Model Organisms in Genetic and Molecular Biology Research Zebra Fish Caenorhabditis elegans Drosophila melanogaster
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Why do We Depend on Animals for Research? To link the physiological appearance with underlying mechanisms Test a gene in its natural environment Provide a model for human disease To test therapies for disease treatment Pharmaceutical Gene therapy To study biochemical and developmental pathways To study a complex interacting system such as immunology
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Naturally occurring mutant animals Animals selected for an induced mutation Transgenic animals Knock-out animals Animals that have “knocked down” genes using RNAi techniques Altering Gene Expression in Animal Models
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The Mouse Advantage Small size, easy to breed Large litter size Short gestation time (19- 21 days) Well established genetics and development Genome fully sequenced (Nature, 2002) Extensive cDNA encyclopedia (Transcriptome) Efficient transgenesis Availability of embryonic stem cell (ES) lines
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What Characterizes a Transgenic Organism? Genome alteration via recombinant DNA-technology Additional genes (“transgenes”) inserted into genome (e.g. EGFP) The transgene is stably integrated into chromosomal DNA The transgene is passed on to the next generation (“goes germline”)
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Nude (Athymic) Mouse is a well-studied animal model of primary immunodeficiency Removing exon 7 of the TGF-7 gene on chromosome 21, Removing exon 7 of the TGF-7 gene on chromosome 21, which codes for thymus growth factor, which codes for thymus growth factor, leads to the  leads to the  absence of a thymus or a vestigial thymus and cell- absence of a thymus or a vestigial thymus and cell- mediated immunodeficiency mediated immunodeficiency nu/nu nu/nu
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Problem in creating transgenic and knock out animals A. The technique is so difficult that only a few labs can do it. A. Embryonic lethality is encountered when the gene regulates animal development.
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Naturally occurring mutant animals Animals selected for an induced mutation Transgenic animals Knock-out animals Animals that have “knocked down” genes using RNAi techniques Altering Gene Expression in Animal Models Caenorhabditis elegans Caenorhabditis elegans
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Life Cycle of C. elegans http://www.wormatlas.org/handbook/anatomyintro/anatomyintro.htm 3 days
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C. elegans has two sexes, hermaphrodite and male. Hermaphrodite- a modified female that in the fourth larval stage makes and stores sperm to be used later to fertilize oocytes produced within the gonad of the same animal after spermatogenesis is finished.
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Lecture 7b - BIO 311 RNAi in C elegans Model Organisms in...

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