Human Genome Engineering using CRISPR/Cas9

Course summary

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 online 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.

Course structure

This introductory course is structured as a combination of lectures and online lab sessions. Our experienced academics and researchers will guide you through generating a knockout and knock in model system using CRISPR/Cas9. In this course we will first introduce you to the general work flow of human genome engineering including the expected timescales to complete a project. We will also discuss the different stages from design of guides to analysis of editing efficiency both in lectures and in the laboratory sessions.

Lectures include:

• Introduction to basics of CRISPR/Cas9 genome engineering and applications
• The Genome Engineering lab work flow of human gene-editing
• Strategies for successful optimal target-specific guide RNA (gRNA) design- discussion of pros and cons of other methods
• DNA repair systems and CRISPR
• Case Study: Living a CRISPR dream – Application of CRISPR in characterization of a  newly identified cancer driver gene
• An opportunity to design your first genome editing experiment

Online hands-on laboratory sessions include:

Workshop A

• Stages in CRISPR genome editing an introduction, an overview of genomic engineering technologies, design of CRISPR-Cas9 reagents
• Guide RNA / Donor design Hands on session - Introduction to guide design strategies, advantages, and limitations

Workshop C

• Verification of trial guide design and plan your first CRISPR editing experiment
• Predictive Genome Engineering strategies

Live streamed sessions from the Genome Engineering Laboratory include:

Workshop B

Transfection of CRISPR components: Preparation of RNP complexes, Electroporation of RNPs and Lipofection of px459 GFP vectors.

Workshop D

Knockout validation Genomic DNA Preparation, Genomic PCR gel analysis of insertions and deletion.

Learning outcomes

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.