67BCAC1Dd01 - University Ghent Faculty of Sciences Academic...

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1 University Ghent Faculty of Sciences Academic Year 2009-2010 Transgenetics of Animal Model Organisms Part Mammals Prof. C. Libert Claude.Libert @ Ugent.be
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2 Contents Part 1: Introduction Chapter 1: The mouse as an experimental animal Chapter 2: wild mice and laboratory mice Chapter 3: Reproduction, breeding and development Chapter 4: The mouse genome Part 2: Mutagenesis Chapter 5. Transgenic animals Overview transgenic animal production Transgenesis in other species Integration of DNA and mosaicism Alternatives to pro-nucleus injection Detection of transgenic animals and expression studies Cryopreservation Intermezzo: situation of transgenesis in farm animals P1, BAC and YAC transgenes and microdissection of chromosomes and transgenesis Inducible transgenesis Production of recombinant proteins in milk of transgenic animals and the environment and transgenesis Promoter research through transgenesis with reporter genes Generate of new cell lines by transgenesis Cell ablation Inhibition of gene expression by transgenic over-expression of anti-sense DNA and by RNAi
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3 Chapter 6. Homologous recombination in ES cells and mutagenesis Definition of ES cells Mouse chimeras derived from ES cells Production of completely ES-derived embryos Homologous Recombination in ES cells Classical way to generate subtle mutations Knockout databases Modification of mitochondrial DNA Importance of genetic background and double knockout mice Expression studies promoters via HR Construction of targeting vectors Diploid mutations in ES cells Knockout sheep and pigs Gene targeting in ES cells using oligonucleotides Chapter 7. Conditional mutagenesis The Cre-LoxP system Non-selectable modifications and removal of selection marker genes Large deletions Gene replacement Chromosomal translocations Targeted integrations and loxP-containing transgenes Conditional modifications Cre transgenic mice
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4 Cell ablation via cre-loxP flp-FRT Problems with cre recombinase Place Specific knock-in of BAC clones RNAi mediated knock-down Shortcuts Combinations of loxP with mutant loxP sequences Knock-in at the ROSA26 locus Chapter 8. Random mutagenesis Introduction Insertion mutagenesis ES Gene-trapping ENU mutagenesis Chapter 9: Large scale mouse mutagenesis programs
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5 ES: Embryonic stem cell DBA: Dilute Brown non-Agouti BALB/c: Baggs Albino TJL: The Jackson Laboratories KO: knock-out NOD: Non Obese Diabetes MRL/lpr: MRL/lymphoproliferation MHC: Major Histocompatibility Complex cM: centi-Morgan TG: transgenic ZP: zona pellucida ICM: inner cell mass OCM: outer cell mass Da: Dalton kb: kilo bases IRES: internal ribosomal entry site BAC: bacterial artificial chromosome ENU: ethyl nitrosurea MMLV: Mouse mammary leukemia virus HSV-tk: herpes simplex virus thymidine kinase Mt-1: metallothioneine PMSG: pregnant mare serum gonadotropine HCG: human chrionotropic gonadotropin CMV: cytomegalovirus EF: elongation factor GH: growth hormone NC: nitrocellulose FSH: follicle stimulating hormone
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This note was uploaded on 05/30/2010 for the course WE CHBIBI0000 taught by Professor Krisvleminckx during the Spring '10 term at Ghent University.

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67BCAC1Dd01 - University Ghent Faculty of Sciences Academic...

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