Genetic Engineering of Mammalian Stem Cells
16-28 February 2015
Wellcome Trust Genome Campus, Hinxton, Cambridge
Application deadline: 10 October 2014
This laboratory-based training course will provide a comprehensive overview and practical laboratory experience of the genetic manipulation of mammalian stem cells.Focusing on mouse ES and human iPS cells, genome informatics, recombineering, CRISPR designer nucleases, transposon technology and gene targeting will be covered in interactive demonstrations, discussions and lectures by international experts. Laboratory work includes the culture and transfection of mouse ES and human iPS cells, design and construction of gene targeting vectors, and the use of PiggyBac transposons and designer nucleases (CRISPRs) for advanced genome engineering and genetic screens. Participants will be trained in the informatics and practical use of public IKMC (International Knockout Mouse Consortium) mouse knockout resources, including the production of mutant mice.Emphasis is placed on the planning/design and successful experimental execution of gene targeting projects and participants should gain the skills necessary to design, construct and target their own genes of interest.
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.
- Professor Francis Stewart (Dresden University of Technology, Dresden, Germany)
- Dr William Skarnes (Wellcome Trust Sanger Institute, Cambridge, UK)
- Dr Pentao Liu (Wellcome Trust Sanger Institute, Cambridge, UK)
- Dr Barry Rosen (Wellcome Trust Sanger Institute, Cambridge, UK)
- Dr Kosuke Yusa (Wellcome Trust Sanger Institute, Cambridge, UK)
2015 Guest speakers (additional speakers TBC shortly)
- Dr David Frendewey (Regeneron Pharmaceuticals, Inc., USA)
- Professor Austin Smith (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK)
- Professor Jin-Soo Kim (Department of Chemistry, Seoul National University, South Korea)
- Dr Ralf Kuhn (Institute for Developmental Genetics, Munich, Germany)
- Dr Wolf Reik (Wellcome Trust Sanger Institute, Cambridge, UK)
- Dr Chad Cowan (Harvard Stem Cell Institute, Massachusetts, USA)
- Professor Nick Hastie (Medical Research Council, UK)
- Professor Christine Mummery (Leiden University Medical Centre, The Netherlands)
- Dr Maria Jasin (Memorial Sloan Kettering Cancer Center, USA)
Feedback from the previous course
“I feel that the course was exceptionally well run and I am extremely grateful to all of the individuals involved that made this possible. The materials provided and the lab work during the course were fantastic and I felt well supported by staff and participants alike.”
“I cannot emphasise enough how much this has transformed my work and scientific confidence.”
“Thank you for everything! The course was a great experience and very useful. Very well organized.”
How to apply
Applicants should be postdoctoral scientists or senior PhD students actively engaged in or soon to commence relevant research. The course is also suitable for facility managers/principal investigators wishing to bring the latest genetic modification technologies into their facilities/groups.
The course tuition fees are subsidised by the Wellcome Trust for scientists based in non-commercial institutions anywhere in the world. This is a residential course and there is a registration fee of £2060 towards board and lodging for non-commercial applicants. The fee for commercial (non-academic) applicants is £4200.
Limited bursaries are available for non-commercial applicants (up to 50% of fee) and are subject to open competition. If you would like to apply for a bursary, please complete the bursary section of the online application form (see below for application process).
Please note: Applications must be supported by a recommendation from a scientific sponsor. This can be your supervisor or head of department. A request for a supporting statement will be sent to your nominated sponsor automatically during the application process. Applicants must ensure that their sponsor provides this supporting statement by the application deadline.
Application deadline: 10 October 2014