Human Genome Engineering using CRISPR/Cas9
CRISPR, one of the powerful scientific stories of the decade, has the potential to revolutionise several areas of biology including medicine and agriculture. Notably CRIPSR is emerging as a rapid diagnostic tool. Due to its simplicity and reproducibility, CRISPR genome engineering could be used successfully to accelerate both basic and applied research in diseases. The Genome Engineering lab at the University of Westminster aims to explore the potential of CRISPR Cas9 technology to combat cancer treatment. We are intensively involved in training Doctoral and graduate students and have successfully applied CRISPR technology to generate several cellular models to study DNA replication dyamics in cancers. The course will provide insights in to the basics of CRISPR gene – editing and hands-on training in human cell engineering using the robust CRISPR Cas9 system. We will also introduce the attendees to successful sgRNA design and analysis strategies and the state-of-art methods that could be used to address their own genome engineering needs. This CRISPR beginners course is aimed at graduate students and scientists with a good understanding of cellular and molecular biology and interest in human genome editing. The course is relevant for researchers venturing into genome engineering for the first time.
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Specific skills you will learn include:
- Knowledge of a simple straight forward work flow to generate CRISPR tools
- Efficient selection and design of guide RNA (gRNA)
- Delivery of CRIPSR tool in human cells, transfection
- Validation of genome editing in engineered cells
Attendees will receive a detailed course guide containing all lecture materials, laboratory protocols and troubleshooting tips.
Timetable as follows:
Human Genome Engineering using CRISPR/Cas9
Day 1: Tuesday 28th April 2020
| 9.00 - 9.45 Arrivals and registration
|
| 9.45-10.00 Welcome and introductions
|
| 10.00-11.00 Inaugural lecture Human Genome Engineering in human disease modelling current applications and limitations in future Dr John J Murphy, Research Director, School of Life Sciences |
| 11.00 -11.30 CRISPR in vitro studies: Working on new cancer driver gene? Case study: My journey with a new cancer breast driver gene and CRISPR/Cas9 genome engineering technology Nadeen Solaiman, Genome Engineering Lab, UoW
|
| 11.30 Tea/Coffee BREAK |
| 12.00 -12.30 AGILENT technologies TALK - Advances in human Genome Engineering Confirmed, waiting for title |
| 112.30 -13.00 Stages in CRISPR genome editing an introduction, an overview of genomic engineering technologies the design of CRISPR-Cas9 reagents Dr Kalpana Surendranath |
| 13.00 -14.00 Lunch BREAK |
| 14.00 - 16.30 Guide RNA / Donor design workshop in computer lab Introduction to: CHOP CHOP, Benchling, Horizon Discovery, Synthego Advantages and limitations. Simple guide cloning methods and successful transfection sytems Dr Kalpana Surendranath
|
| 16.30 – 17.00 Tea/Coffee BREAK |
|
17.00-19.00 Genome Engineering Laboratory C5.02 Lab session:1 Work with cells to be transfected tomorrow, assessing cell count and viability () followed by plating cells for transfection |
| END of DAY1 |
Human Genome Engineering using CRISPR/Cas9
Day 2: Wednesday 29th April 2020
| 9.30 - 10.00 CRISPR-Cas9 genome editing and the DNA damage response Dr Kanagaraj Radhakrishnan (Francis Crick Institute)
|
| 10.00 – 10.30 THERMOFISHER TALK-Advances in human Genome Engineering Confirmed waiting for title |
| 10.30 – 11.00 BREAK |
| 11.00 - 13.00 Lab session:2 Transfection of CRISPR components: Preparation of RNP complexes, Electroporation of RNPs and lipofection of CRISPR mcherry vectors |
| 13.00 - 14.00 Lunch Break |
| 14.00 - 14.30 Cas9 enzyme purification and applications |
| 14.30 - 15.00 Advances in human Genome Engineering VectorBuilder: CRISPR Vector and Virus solutions Matthew Wheeler, Associate Director Vector Builder GmbH |
| 15.00 - 15.30 Coffee break/ networking |
| 15.30-18.00 Lab session:3 Setting up RNP transfection Verification of trial guide design and plan for your first CRISPR editing experiment |
| 19.00 -21.00 Dinner – venue |
Human Genome Engineering using CRISPR/Cas9
Day 3: Thursday 30th April 2020
| 9.30-12.00 Lab session:4 Examine mcherry positive cells via microscopy |
| 12.00 – 13.00 Lunch Break |
| 13.00- 14. 00 Advances in human Genome Engineering Genome Editing: A New Era in Molecular Biology Marc Davies, Account Manager, New England, Biolabs, UK |
| 14.00 – 14.30 Purification of Cas9 protein: applications |
| 14.30-15.30 Verification of genome editing, bioinformatic tools and workflow |
| 15.30 – 16.00 Summary of the workshop Guidance and Feedback |
| 16.00 - 16.30 Closing remarks and acknowledgements |
Event contact details
Dr Kalpana Surendranath Email: k.surendranath1@westminster.ac.uk Telephone: 020 7911 5000 ext. 64121 Event website
Location
School of Life Sciences
University of Westminster
115 New Cavendish Street
London
W1W 6UW