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Cancer Research (Subject) / V.9 Mouse models (Lesson)
- What are the uses of CreLoxP and CRISPR Cas technologies? Cre-LoxP technology allows for tissue-specific gene activation or inactivation. CRISPR-Cas9 technology enables us to precisely edit genomes with a plethora of applications in biology, biomedicine and crop science
- What does genome engineering / gene editing describe? It describes the targeted manipulation of the genetic make-up at the oocyte stage with the aim to stably introduce an inheritable alteration of the genotype.
- What is the difference of Genome engineering and forward genetic screens? Genome engineering: targeted manipulation to activate or deactivate specific genetic elements Forward genetic screens: phenotype-driven analyses of undirected mutagenesis (f.e. targeted breeding)
- Ordnen Sie die folgenden experimentellen Vorhaben den entsprechenden Methoden zu a. Zebrafisch mit Photorezeptor Degenration b. Korrektur eines Aminosärenaustauschs im Genom c. Gewebespezifische Aktivierung eines Gens 1. Cre-LoxP 2. chemische Mutagenzien 3. CRISPR/Cas9 a. Zebrafisch mit Photorezeptor Degenration --> 2. chemische Mutagenzien b. Korrektur eines Aminosärenaustauschs im Genom --> 3. CRISPR/Cas9 c. Gewebespezifische Aktivierung eines Gens --> 1. Cre-LoxP
- Which are characteristics of different model systems? -Gravidity time -Generation time -No. of offsprings-Versatility of models -Housing space needed -Organ development
- What is the effect of ENU (=N-ethyl-N-nitrosourea)? -alkylating agent-mainly causes A->T and AT->GC transitions-targets spermatogonial stem cells
- What are forward genetic screens used for? What are the model organisms? Forward genetic screens can be performed to identify phenotypes and their underlying genes by undirected mutagenesis. Main model organisms are mice or zebrafish
- How to generate a transgenic mouse? a. „Quick and dirty“: microinjection of DNA into pronuclei b. Targeted insertion by recombination into homologous loci or todefined genomic regions
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