Laboratory

Genetic Engineering of Mammalian Stem Cells

April 11th - 23rd 2016

Wellcome Genome Campus, Hinxton, UK

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 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.

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!”

Programme

The programme will include lecture and practical
laboratory/computer-based sessions covering the following topics:

1. Informatics
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

Instructors and speakers

Course instructors
Francis Stewart Dresden University of Technology,
Dresden, Germany
William Skarnes Wellcome Trust Sanger Institute,
Cambridge, UK
Pentao Liu Wellcome Trust Sanger Institute, Cambridge,
UK
Barry Rosen AstraZeneca, Cambridge, UK
Kosuke Yusa Wellcome Trust Sanger Institute, Cambridge,
UK
Bon-Kyoung Koo Wellcome Trust – MRC Cambridge Stem Cell
Institute, Cambridge, UK

Confirmed guest speakers
Jacob Corn University of California, Berkeley,
USA
David Frendewey
Regeneron Pharmaceuticals, Inc., USA
Austin Smith Wellcome Trust Centre for Stem Cell
Research, Cambridge, UK
Kevin Eggan Harvard Stem Cell Institute, Massachusetts,
USA
Nick Hastie Medical Research Council, UK
Christine Mummery Leiden University Medical Centre, The
Netherlands
Jennifer Nichols Wellcome Trust Centre for Stem Cell
Research, Cambridge, UK
Allan Bradley Wellcome Trust Sanger Institute,
Cambridge, UK
Gavin Kelsey Babraham Institute, UK
Martin Jinek
University of Zurich, Switzerland

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 is subsidised by the Wellcome Genome Campus Advanced Courses
and Scientific Conferences Programme. This is a residential
course and
there is a fee of £2100 towards board and lodging for non-commercial
applicants. Please contact us for the commercial fee.

Applications
Applications for this course can now be completed online. If you have any
problems with the online application process, please contact us.

Please note: Applications
must be supported by a
recommendation from a scientific or clinical sponsor (e.g. 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. Applications without a supporting
statement cannot be considered.

Deadlines
Deadline for Applications: Closed

Bursaries

Bursaries
Advanced Courses are subsidised for non-commercial applicants from
anywhere in the world. Limited bursaries are
available (up to 50% of the
course fee) and are awarded on merit. If you would like to apply for a
bursary, please complete the bursary section of the online application
form.

Please note that both the applicant
and sponsor are required to provide
a justification for the
bursary as part of the application.

Bursary terms and conditions

UK Courses (held at the Wellcome Genome Campus, Hinxton,
Cambridge)
A
limited number of bursaries are available for each course. These are
awarded by the selection committee according to merit. The bursary
covers a maximum of 50% 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 to attend UK
courses.

Overseas Courses (held outside of the UK)
A
limited number of bursaries are available for each course. These are
awarded on merit to cover travel, accommodation and sustenance. The
maximum award for travel (economy class) will be £750.

Bursaries can be applied for as part of the course application form.
Applicants
will be notified of a bursary award along with their place on
the
course, usually within one month of the application deadline. The
decision of the selection committee is
final.

Back to top Back to top