Genetic Engineering of Mammalian Stem Cells - Training Course

 

Course - Cambridge, United Kingdom

 

Course programme

1. Informatics
The informatics underlying the visualization and interpretation of gene structures will be presented focusing on the practical design of gene targeting strategies. Students will also develop their own CRISPR-mediated gene targeting designs using web-based tools and learn how to exploit the public IKMC/IMPC mouse gene targeting/phenotyping resources.

2. Recombineering of BACs and Gene Targeting Vectors
Students will use recombineering to make modified BAC transgenes and gene targeting constructs. The theoretical principles underlying both recombineering and rational targeting vector design will be presented in lectures and practical exercises.

3. Modular Gene Targeting Vectors
Students will learn to apply modular approaches to vector assembly and gene targeting strategies.Students will build their own vectors for CRISPR-based gene targeting using Gibson assembly.

4. Stem Cell Culture/Gene Targeting
Students will learn feeder-free culture of mouse ES cells and human iPS cells including a number of standard transfection techniques (electroporation, nucleofection, lipofection). Transfection, selection, picking, processing and genotyping for gene targeting in ES cells will be covered.

5. CRISPR Designer Nucleases
A CRISPR mediated homologous recombination experiment will be performed in human IPS cells.Practical application of the CRISPR system to undertake in vitro genetic screens will also be explored.

6. Transposon Technology/IPS Cell Generation
Students will be introduced to the uses of the highly efficient PiggyBac transposon systems for BAC transgenesis and reprogramming for iPS.

 

Organization: Wellcome Trust

 

 

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