Introductory Genome Engineering Workshop
General context
After organizing several successful symposia on genome engineering and CRISPR in the last years, VIB Training now offers a hands-on genome engineering workshop. After an introduction to CRISPR (history, biology and applications), we will focus on two experiments:
- NHEJ experiment in maize
- HDR editing experiment in yeast
NHEJ experiment:
- Designing amplicons before conducting experiment and general QC
- gRNA design (design rules, efficiency, predicting indel outcomes...)
- Performing and analyzing genotyping results with Sanger-based approaches
- Off-target analysis
HDR editing experiment:
- gRNA design, choice and rationale, importance of PAM distance in HR editing
- donor design
- predictions don’t always reflect reality , understand different reasons for variable efficiencies between gRNAs
- learn about challenges of HDR editing and different ways to surmount them
The training will be a combination of lectures by internal experts, practicals and discussion on the obtained results. There will be a full day introduction followed by 3 half-a-day sessions. The sessions will take place in Leuven and in Ghent (alternating), and in between there will be some assignments for the participants (genotyping, summarize results, reading assigned articles and preparing short presentations in groups). We estimate that the additional amount of work outside of the course will be 8 hours.
Note that if you register, we expect you to participate in all 4 sessions.
PhD students, technicians and postdocs interested in learning more on the practical aspects of genome engineering. Preferably those with little or no experience in CRISPR.
Introduction to Benchling and Geneious.
- This course is free for VIB participants
- Registration fee for non-VIB academics is 200 euro including VAT.
- This course is not open for industry.
- Lunch will be provided.
- Note that upon no show without valid justification you will be blacklisted for the VIB training program for 1 year. Click here for more information.
Trainers
Sibylle Vonesch
Thomas Jacobs
Program
Perform a NHEJ experiment – part 1 (7 February 2023, 10h - 17h, Leuven)
Design a gRNA and target site for knock-out of maize genes.
Assignment:
- Test genotyping primers and propose gRNAs for KO
- Genotype supplied sample DNA and bring results to class
Perform a NHEJ experiment – part 2 (13 March 2023, 10h - 14h, Ghent)
Evaluation of sequencing results.
Assignment: Summarize results and prepare report on indel efficiency and off-target effects.
HDR editing experiment – part 1 (21 April 2023, 10h -14h, Leuven)
Insertion of PTCs and missense alleles via single nucleotide changes in the yeast ADE2 gene:
- gRNA design for HDR mediated precision editing, efficiency and off-target potential
- importance of PAM distance for HR editing, alternative Cas nucleases
- donor design
Assignment: Read assigned HDR boosting article and prepare short presentation in groups.
HDR editing experiment – part 2 (8 May 2023, 10h - 14h, Ghent)
Determine efficiency (visual read-outs and sequencing results) with different guides/target sites and compare.
Comparison predicted/obtained efficiencies, reasons for variable efficiencies between guides.
Discussion on strategies to boost HDR
Practical info
07 February 2023 - 21 April 2023
Leuven - Park Inn by Radisson
Martelarenlaan 36
3010 Leuven
Belgium
13 March 2023 - 08 May 2023
Ghent - VIB/UGent FSVM II
Technologiepark 75
9052 Zwijnaarde
Belgium
07 February 2023 - 21 April 2023
Leuven - Park Inn by Radisson
The hotel is located 300m from Leuven Central Station. The hotel connects to Leuven Central Station by a pedestrian bridge.
13 March 2023 - 08 May 2023
Ghent - VIB/UGent FSVM II
From Ghent Sint-Pieters station, you can take bus 49, 50 or 70 to Technologiepark. Please check Routeplanner De Lijn for schedules.
07 February 2023 - 21 April 2023
Leuven - Park Inn by Radisson
Park Inn hotel has no own parking. It is possible to park underground in P1-Parking Station Leuven, Martelarenlaan 4, 3010 Leuven or in parking De Bond, Martelarenlaan 18, 3010 Leuven.
13 March 2023 - 08 May 2023
Ghent - VIB/UGent FSVM II
There is only one entrance to Technologiepark. At the entrance, please take a ticket and have it validated at the reception desk of the FSVM building. Parking will only be allowed in regular parking spots (for instance in front of the building) and in the new parking tower. Parking alongside the roads or in other places where there is no regular parking is prohibited and after initial warnings, fines will be issued (€ 50).
07 February 2023 - 21 April 2023
Leuven - Park Inn by Radisson
+32 16 61 66 02
sales.leuven@parkinn.com