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Genetic Engineering of Mammalian Stem Cells

10-22 March 2014

Wellcome Trust Genome Campus, Hinxton, Cambridge

Now closed for applications. Please email to join our mailing list.

Course summary

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, recent advances in genome informatics, recombineering, custom nucleases (CRISPRs/TALENs), transposon technology and conditional gene targeting will be covered in lectures and through interactive demonstrations. Laboratory work includes the culture and transfection of ES cells, design and construction of gene targeting vectors by BAC recombineering, and the use of PiggyBac transposons and designer nucleases (CRISPRs/TALENs) for advanced genome engineering. Participants will be trained in the informatics and practical use of public gene targeting resources, including the production of mutant mice from the IKMC (International Knockout Mouse Consortium) resource. Emphasis is placed on the planning/design and successful experimental execution of gene targeting projects.

Course programme

1. Informatics
The informatics underlying the visualization of gene structures and the design of gene targeting vectors, recombineering oligos and genotyping primers will be demonstrated. Students will also run their own gene targeting designs using web-based tools and learn how to use the public IKMC gene targeting resources.

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

3. Stem Cell Culture
Students will learn feeder-free culture of ES cells derived from 129 and C57BL/6 mouse strains and human iPS cells.ES cell colonies will be picked, expanded and frozen and subsequently thawed to test their viability.

4. Gene Targeting in mouse ES Cells
Transfection, selection, picking, processing and genotyping for gene targeting in ES cells will be covered.

5. Transposon Technology
Students will be introduced to the uses of the highly efficient piggyBac transposon systems for BAC transgenesis, insertional mutagenesis and reprogramming for iPS.

6. Site Specific Nucleases (TALENs/CRISPRs)
Students will transfect ES cells with TALEN/CRISPRs and learn about the applications of custom site-specific nucleases in stem cell genome engineering.

7. Modular Targeting Resources
Students will assemble a variety of modular knock-in targeting vectors from IKMC resources and analyse their integrity.Recombination Mediated Cassette Exchange (RMCE) using Flp and Cre recombinases will be used to modify IKMC conditional alleles directly in ES cells.

8. Homozygous ES Cells
Students will learn about approaches used to generate homozygous ES cells and the analysis of cellular phenotypes.

Course instructors

Guest speakers

  • Professor Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK)
  • Dr David Frendewey (Regeneron Pharmaceuticals, Inc., USA)
  • Dr Timm Schroeder (Helmholtz Zentrum München, Germany)
  • Professor Austin Smith (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK)
  • Dr Anton Wutz (Department of Biochemistry, University of Cambridge, UK)
  • Professor Jin-Soo Kim (Department of Chemistry, Seoul National University, South Korea)
  • Dr Ralf Kuhn (Institute for Developmental Genetics, Munich, Germany)
  • Dr Ludovic Vallier (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK)
  • Dr Wolf Reik (Wellcome Trust Sanger Institute, Cambridge, UK)
  • Dr Chad Cowan (Harvard Stem Cell Institute, Massachusetts, USA)
  • Dr Kosuke Yusa (Wellcome Trust Sanger Institute, Cambridge, UK)

Feedback from the previous course

“I would like to express my gratitude to all course assistants and organizers for making such a complex and well-working course possible. It was a real pleasure being here and I will definitely apply lots of the new things I learned into my current work flow, and I am sure it will bring lots of improvements and new strategies. THANKS a LOT”

“Very grateful for the excellent organization and the topics covered, as well as for the availability and enthusiasm of both, the course instructors and the speakers.”

“Thank you for giving me the opportunity to attend the course. I've learned a lot and met some amazing people”

How to apply

Prerequisites
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.

Cost
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 £1960 towards board and lodging for non-commercial applicants. The fee for commercial (non-academic) applicants is £4000.

Bursaries
Limited bursaries are available for non-commercial applicants (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).

Deadline
Now closed for applications. Please email to join our mailing list.

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