Genetic Manipulation of ES Cells

Course summary
This course will provide a comprehensive overview and practical laboratory experience of the genetic manipulation of ES cells for a broad range of applications. Recent advances in genome informatics, BAC recombineering, transposon technology and conditional gene targeting will be covered in theoretical sessions and demonstrations.Laboratory work will focus on the culture and transfection of ES cells, construction of gene targeting vectors, characterization of random gene trap insertions and genotyping of gene targeting events. The course is subsidised for academic participants and limited bursaries are available through funding provided by The Wellcome Trust and the European Conditional Mouse Mutagenesis Program ( EUCOMM). See below for further information.

Course programme
1. Recombineering of Conditional Gene Targeting Vectors
Students will construct conditional gene targeting constructs from BACs using recombineering technology. Participants will be invited to nominate genes of interest ahead of the course, which the instructors will incorporate into the practical programme where possible.

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

3. Gene Trap Mutagenesis
Students will co-electroporate ES cells with a transposase expression plasmid and transposon-based gene trap construct. ES cell colonies will be picked and expanded for the preparation of genomic DNA. Splinkerette PCR will be performed to amplify and sequence genomic DNA flanking the insertion site.

4. Conditional Gene Targeting
Students will electroporate ES cells with promoterless and promoter-driven gene targeting constructs. ES cell colonies will be picked and expanded for the preparation of genomic DNA.Long-range PCR will be performed to identify correctly targeted events.

5. Informatics
The informatics necessary for the design of gene targeting vectors, recombineering oligos and genotyping primers will be demonstrated. Students will run their own designs using a web-based design tool.Methods for mapping splinkerette sequence tags to the mouse genome will be demonstrated and students will analyse and annotate sequence reads from random gene trap insertions.

Course instructors
Professor Allan Bradley (Wellcome Trust Sanger Institute, Cambridge)
Professor Francis Stewart (Dresden University of Technology, Dresden)
Dr William Skarnes (Wellcome Trust Sanger institute, Cambridge)
Dr Pentao Liu (Wellcome Trust Sanger Institute, Cambridge)
Dr Barry Rosen (Wellcome Trust Sanger Institute, Cambridge)

Guest speakers
Andras Nagy (Samuel Lunenfeld Research Institute, Toronto, Canada)
Andrew Smith (University of Edinburgh, UK)
Austin Smith (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK)
Drew Murphy (Regeneron, NY, USA)
Eugene Yu (Roswell Park Cancer Institute, NY, USA)
Junji Takeda (Osaka University, Japan)

Cost
The course tuition fees are subsidised by the Wellcome Trust for scientists based in academic institutions anywhere in the world. This is a residential course, without exception, and there is a registration fee of £1960 towards board and lodging for academic applicants. The fee for commercial (non-academic) applicants is £7500.

Bursaries
Limited bursaries are available for academic applicants (50 per cent of fee) and are subject to open competition. Funding for bursaries is provided by The Wellcome Trust and the European Conditional Mouse Mutagenesis Program ( EUCOMM).

Applications
Applicants will be required to complete an application form containing their CV together with a 300-word outline of the relevance of the course to their research. Please note that documentation supporting their application will be required from the applicant's supervisor/head of department.

Bursaries for Wellcome Trust Advanced Courses
The Wellcome Trust subsidises all courses and workshops, so there are no tuition fees. The ‘course fee' for the UK courses (held at Hinxton) covers on-site accommodation and food for the duration of the course. Bursaries are available to assist with UK course fees, but not travel costs. We are committed to building scientific capacity in developing countries and are expanding the number of courses and workshops run outside of the UK (‘overseas' courses). There is no fee for these courses and bursaries are available for travel, sustenance and accommodation. Please see below for full details.

UK Courses (held at the Wellcome Trust Genome Campus in Hinxton)
A limited number of bursaries are available for each course. These are awarded by the selection committee according to merit. The bursary usually covers 50 per cent of the course fee, though in exceptional circumstances an application for the total course fee may be considered. Where there are many bursary applications, the selection committee may issue smaller amounts. We cannot assist with travel costs. All bursary requests to attend UK courses must be accompanied by a bursary application form. This must be completed and signed by your scientific sponsor (supervisor or head of department) and include justification for the bursary. Applicants will be notified of a bursary award along with their place on the course, usually within one month of the application deadline. We will reject incomplete applications. Both the application form and sponsor's letter must be received before the application deadline. The decision of the selection committee is final.

Overseas courses (held outside the UK)
A number of bursaries are available for each overseas course. These are awarded by the selection committee according to merit. There is no course fee.The bursary will cover travel, accommodation and sustenance. The maximum award for travel (in economy/second class) will be £750. All likely travel costs must be detailed in the overseas bursary application form, which must accompany all bursary requests for courses held outside of the UK. We will notify you of the chosen hotel and sustenance award if successful. The selection committee may judge that a partial award should be granted. We will reject incomplete applications. Both the application form and sponsor's letter must be received before the application deadline.