Genetic Engineering of Mammalian Stem Cells

This popular 2-week laboratory-based training course provides a
comprehensive overview and practical laboratory experience of the genetic
manipulation of mammalian stem cells, focusing on human iPS cells.

The intensive, hands-on programme covers cutting-edge techniques in this
exciting and fast-moving field, including genome informatics, vector
construction employing recombineering and synthetic biology methods, and
genome editing strategies using CRISPR-Cas9. The course also includes
interactive demonstrations, discussions and lectures by international
experts.

Laboratory work includes the use of 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 course is aimed at 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.

Feedback from the 2016 course

• “The course was excellent, very well-run and planned and highly useful to my current and planned projects. I appreciated the access and engagement with instructors, speakers and coursemates, which provided multiple avenues for learning and discussion. I enjoyed the intellectual experience, and appreciate the time and efforts of the instructors, course organisers and administrative and laboratory staff.”
• “This is one of the best courses I have visited so far.”
• “Overall the course was of a very high standard with all the instructors doing a great job and their time and effort is really appreciated.”
• “Overall the quality and standard of the course was excellent. The team of instructors/organizers was absolutely fantastic and the panel of speakers is great.”
• “I very much enjoyed the course and have learned a lot. I would like to thank the organisers for putting in all this effort to organise a fantastic course.”

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

Informatics
The visualization and interpretation of gene models will be presented
focusing on the practical design of gene targeting strategies. Students
will also have the opportunity to develop their own CRISPR-mediated gene
targeting designs using web-based tools and learn how to exploit public
genomic data resources.

Modular Gene Targeting Vectors
Students will learn to apply modular approaches to vector assembly and
gene targeting strategies and engage in the construction and analysis of
vectors for CRISPR-assisted gene targeting.

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 alleles. Cell manipulation techniques including picking,
expansion, archiving and genotyping of iPS cell clones will also be
covered.

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.

iPS Cell Generation
Students will be introduced to protocols for highly efficient
reprogramming of mouse and human somatic cells to induced pluripotent
stem cells.

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

Learning Outcomes
After attending this course, participants will be able to:

• Assess targeting strategies within a genomic context, identifying suitable targets and CRISPR reagents
• Design and build vectors for CRISPR-assisted gene targeting using modular construction approaches
• Apply cellular reprogramming methods to derive induced pluripotent stem cells for human and mouse
• Undertake genome editing of human iPS cells, including manipulation of cells under culture conditions, applying different transfection techniques
• Develop genome-wide screens exploiting lentiviral transduction methods, including analysis using next-generation sequencing
• Investigate new research techniques, exploiting tissue-derived organoids to explore gene function and cellular interactions

Course instructors
Andrew Bassett Wellcome Trust Sanger Institute, UK
Bon-Kyoung
Koo Wellcome Trust – MRC Cambridge Stem Cell Institute,
UK
Pentao
Liu Wellcome Trust Sanger Institute, UK
Barry Rosen AstraZeneca, Cambridge, UK
Mark
Thomas Wellcome Trust Sanger Institute, UK
Kosuke
Yusa Wellcome Trust Sanger Institute, UK

Guest speakers
Chad Cowan
Harvard Stem Cell Institute, USA
Hans Clevers Hubrecht
Institute, The Netherlands
Frank
Buchholz University of Dresden, Germany
David Frendewey Regeneron, USA
Christine
Mummery Leiden University Medical Center, The Netherlands
Kathy
Niakan
Francis Crick Institute, UK
Jennifer
Nichols
Wellcome Trust Centre for Stem Cell Research, UK
Wolf
Reik Babraham Institute, UK

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 £2140 towards
board and lodging for non-commercial applicants. Please contact us for
the commercial fee.

Additional limited bursaries are available (up to 50%
of the course fee)
and are awarded on merit. Please see the “Bursaries”
tab for details.

Applications
Applications for this course can 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

Travel visas
Please contact the
event organiser if you require a letter to support a
travel visa
application. Note that letters will be provided to confirmed
attendees.

Non-European Economic Area or Swiss nationals may be required to have a
visa to enter the UK.
Early application is strongly advised, as this process can take 6-8 weeks
or longer.

Please visit the following websites for further information:
UK Border Agency website and information for general visitors and business
visitors.

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 £2140 towards
board and lodging for non-commercial applicants. Please contact us for
the commercial fee.

Bursaries
Advanced Courses are subsidised for non-commercial
applicants from anywhere in the world. Additional, 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.