COURSE REPORT

Course Name: 3rd Programmable Nucleases (CRISPR/Cas9) Transgenesis Course

Course Dates: 16 April - 20 April 2018

Location: Czech Center for Phenogenomics, Institute of Molecular Genetics AS CR,

Prumylsova 595, Vestec, Czech Republic

Course Coordinator/Point of Contact: Assoc. Prof. Radislav Sedlacek, PhD

Number of Students: 12

Tutors:

Bernd Zetsche Broad Institute, USA

Lluis Montoliu Centro Nacional de Biotecnologia (CNB-CSIC), Spain

Brandon Willis UC Davis, USA

Astrid Gall Ensembl, UK

Ralf Kuehn Max Delbrück Center for Molecular Medicine, Germany

Gurumurthy Channabasavaiah University of Nebraska Medical Center, USA

Steve Riedmuller Synthetic Genomics, Inc., USA

Marie-Christine Birling ICS, Phenomin, France

Guillaume Pavlovic ICS, Phenomin, France

Petr Kasparek IMG, Prague

Lectures:

Genome editing with programmable nucleases: general overview (Lluis Montoliu)

Ensembel session (Astrid Gall)

Rapid DNA Synthesis for CRISPR applications; Leveraging the BioXp for your Gene editing workflow (Steve Riedmuller)

Design and preparation of gRNA (Brandon Willis)

Designing your best RNA guides for CRISPR experiments: multiple solutions available (Lluis Montoliu)

Electroporation of zygotes (Ralf Kuhn)

Design of targeting vectors (Brandon Willis)

Easi-CRISPR and GONAD: CRISPRing Mouse Genome Made Easy (Gurumurthy Channabasavaiah)

CRISPR for the generation of innovative rodent models (Marie-Christine Birling)

Genome editing with Cpf1 (Bernd Zetsche)

Analyzing the outcome of genome editing: screening, genotyping and quality control (Guillaume Pavlovic)

General Lecture: From genotype to phenotype (Guillaume Pavlovic)

Practical training sessions/Individual projects:

Zygote electroporation and Digital PCR – Irena Jenickova, Jana Kopkanova (IMG/CCP),

Guillaume Pavlovic

Individual projects – Brandon Willis, Marie- Christine Birling, Lluis Montoliu, Bernd Zetsche,

Petr Kasparek

Student´s evaluation of the course:

The course has been evaluated as Good and Excellent, with prevalence at Excellent.

Summary:

The course has been designed to give practical, hands-on training in current technologies and

methods employing programmable nucleases (CRISPR/Cas9) - mediated genome manipulations

in the generation of mice mutants.

Participants gained theoretical and practical knowledge from our expert instructors and had the opportunity for expert insight for their own individual project.

Participants learned how to:

• Plan and choose the targeting strategy • Apply the targeting tools and validate CRISPR activity • Deliver targeting tools into zygotes • Identify CRISPR induced genomic modifications

Date: 15 May 2018