Genetic Engineering of Mammalian Stem Cells
11-23 April 2016
Wellcome Trust Genome Campus, Hinxton, Cambridge
This laboratory-based training course will provide a comprehensive overview and practical laboratory experience of the genetic manipulation of mammalian stem cells, focusing on human iPS cells.Strategies for genome engineering of mouse models and mutant mouse resources produced by the International Mouse Phenotyping Consortium (IMPC) will also be discussed.Genome informatics, vector construction employing recombineering and synthetic biology methods, and genome editing strategies using CRISPR-Cas9 will be covered in interactive demonstrations, discussions and lectures by international experts.
Laboratory work includes the use CRISPR-Cas9 for advanced genome engineering and genetic screens, the design and construction of gene targeting vectors, the culture and transfection of human iPS cells and the derivation of iPS cells and organoid stem cells. An emphasis will be 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 visualization and interpretation of gene models 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 IMPC mouse gene targeting/phenotyping resources.
2. Recombineering of BACs and Gene Targeting Vectors
Participants 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 and engage in designing and building their own vectors for CRISPR-assisted gene targeting.
4. Stem Cell Culture/Genome Editing
Genome editing experiments will be performed where students will learn feeder-free culture of human iPS cells and transfection techniques for plasmid- and protein-based delivery of Cas9 to engineer a variety of useful allelesPicking, expansion, archiving and genotyping of iPS cell clones will also be covered.
5. Genome-wide screens
Practical application of the CRISPR system to undertake in vitro genetic screens will also be explored. This includes the design of genome-wide screens, lentiviral transduction of mouse ES cells and bioinformatic analysis of next-generation sequencing data.
6. iPS Cell Generation
Students will be introduced to protocols for highly efficient reprogramming of mouse and human somatic cells to induced pluripotent stem cells.
7. Organoid derivation and differentiation
Organoids (primary 3D epithelial culture) can be derived from many tissues of any mouse strain and enables us to explore gene functions in diverse types of primary cells. A protocol for the establishment of murine intestinal organoids will be introduced. Several transgenic organoids will be shown as examples
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 (AstraZeneca, Cambridge, UK)
Dr Kosuke Yusa (Wellcome Trust Sanger Institute, Cambridge, UK)
Dr Bon-Kyoung Koo (Wellcome Trust – MRC Cambridge Stem Cell Institute, Cambridge, UK)
Confirmed 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)
Dr Kevin Eggan (Harvard Stem Cell Institute, Massachusetts, USA)
Professor Nick Hastie (Medical Research Council, UK)
Professor Christine Mummery (Leiden University Medical Centre, The Netherlands)
Dr Jennifer Nichols (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK)
Professor Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK)
Dr Gavin Kelsey (Babraham Institute, UK)
Feedback from the 2015 course
“The quality and attitude of the instructors was world-class; they created a top tier course and delivered state-of-the-art information. I would highly recommend this Advanced Course to anyone seeking to perform genetic manipulations in mammalian cells. My deepest thanks to all those involved in putting this together.”
“This was an amazing course! It covered a wide range of state of the art topics and the practical parts were perfect to get in close contact to these experts in their fields. The motivation and performance of all teachers and lecturers was outstanding and encouraging! I want to thank all the organizers, teachers, lecturers and participants for this great experience.”
“This is an awesome course! The instructors are amazing and their dedication and commitment in organizing such a course every year is commendable. The selection of course participants is also excellent. Wonderful all around.”
“This course is amazing! Thank you to all the wonderful instructors!”
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 researchers based in non-commercial institutions anywhere in the world. This is a residential course and there is a fee of £2100 towards board and lodging for non-commercial applicants. Commercial applicants should contact us for the commercial fee.
Limited bursaries are available for non-commercial applicants (up to 50 per cent 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.
Deadline for applications: 6 November 2016