(SW5) Genome Engineering Using CRISPR/Cas9
On behalf of the ABRF Executive Board and ABRF Education Committee, we welcome you to the second annual ABRF Satellite Workshop on Genome Editing. While we will adhere to a similar format as last year (based on the positive feedback from these participants), we have added one significant new feature. We would like to encourage participants, who register for the workshop, to present their research problems to the whole group or to request guidance on how best to utilize genome editing technologies to address a question within their project. We call this part of the session, Interactive Learning. We are incorporating this new session because the field of gene editing is rapidly evolving and questions regarding ease-of-use, reproducibility and robustness abound. We will provide a tutorial on gene editing, enable significant interactions among all participants and discuss the current state of this technology as a research tool and as it relates to biomedical core function. We are looking forward to expanding the breath of this workshop by discussing gene editing applications on a case-by-case basis.
Eric B. Kmiec & CB Gurumurthy
Kym Delventhal, Stowers Institute
Gerald Marsischky,Harvard University
Timothy Dalham,University of Utah
1. Offer participants an overall understanding of CRISPR/Cas9 technology and the applications enhanced by use of the system.
2. How to design experiments using either CRISPR alone or CRISPR with homologous donor recombination
3. How to screen for targeted mutations
4. Tools available and applied applications to facilitate gene editing
5. Provide insight on best practices of offering this service through a core facility
The field of genome engineering has improved drastically in the past four years. Currently the CRISPR/Cas9 system is the most popular system due to its ease of use and high levels of success rates. Scientific researchers are now equipped with a system that has the potential to make any desired change in the genome. While building a CRISPR construct is straightforward to do, many researchers are just beginning to utilize this technology in their projects and the other requirements may seem daunting. Core facilities that offer genome engineering services provide assistance in managing the overall project and offer advice on how best to design the experiment to ensure success.
Core directors, managers and technicians that desire an understanding of the CRISPR/Cas9 system in order to support genome engineering services at their organization or to collaborate on projects using these technologies.
CRISPR/Cas9 basics, definitions of project types (knockout, transgenic), lingo introduced
Designing CRISPR/Cas9 target sites
Designing homologous donor recombination construct
Delivery of genome editing components (cells, mouse, zebrafish, drosophila, c. elegans, etc.)
Screening for expected mutations (PCR, T7 endonuclease I digest, Sanger sequencing, droplet digital PCR, HRMA)
Internet connection capability for all attendees. Typical AV equipment such as LCD projector and screen. Attendees will be required to bring their own laptop computer.
Part 1: The Foundation of CRISPR/Cas9 Genome Editing
8:30am - 8:50am: Welcome and Remarks from the Moderator.
Eric Kmiec, Gene Editing Institute, Helen F Graham Cancer Center and Research Institute , University of Delaware.
8:50am – 9:20am: Genome engineering using the CRISPR/Cas9 system; basics and recent trends
CB Gurumurthy, University of Nebraska Medical Center.
Part 2: Experimental Design and Identifying CRISPR mutations
9:20am – 9:50am: Genome Engineering Services and operations; a core facility perspective
Kym Delventhal, Stowers Institute.
10:00am – 10:30am: HDR considerations and a few examples of designing CRISPR animal models
CB Gurumurthy, University of Nebraska Medical Center.
10:30am- 10:45am Break
Part 3: Interactive Learning and Short Presentations.
10:45am – 12:00pm:
This session will include a brief 2-3 slide (5 minute) talk from any workshop participant who wishes to discuss their particular project and wherein they are considering using genome editing. The speaker panel will try to provide guidance and suggestions regarding the feasibility of the project and suggest the best approach. This session is intended to facilitate communication among speakers and participants in the workshop and to provide technological advice for gene editing approaches in biomedical research.
* Presentation by workshop participants is totally voluntary.
12:00pm – 1:00pm: Lunch
Part 4: Experimental Design and Identifying CRISPR mutations Continued
1:30pm – 2.00pm: Screening for Engineered Genome Alterations.
Gerald Marsischky, Harvard University.
Part 5: Commercial tools for Gene Editing
2:00pm – 2:30pm: Improved fidelity of genome editing by the IDT Alt-R CRISPR-Cas9 and Cpf1 systems
Christopher Vakulskas, Integrated DNA Technologies
2:30pm – 3:00pm: A Highly Versatile Tool for CRISPR Screening
Christian H. Lytle, Molecular Biology Core Facility Manager, Geisel School of Medicine at Dartmouth
Part 6: The Future of Genome Editing
3:00pm - 3:30pm: The future of University CRISPR-Cas9 Cores
Timothy Dalham, University of Utah.
3:30pm – 3:45pm Break
Part 7: Panel discussion on the Future of Gene Editing Cores
Eric B. Kmiec, Moderator , Gene Editing Institute, Helen F Graham Cancer Center and Research Institute , University of Delaware.
The formal workshop will close with a general discussion including speakers and participants regarding the future of gene editing technologies and how best to incorporate them into a biomedical research core facility.
Attendees will be expected to bring their own laptops. No special software is needed other than the ability to connect to the Internet.