Basics of CRISPR/Cas with hands-on techniques

In this course, we will cover the basics of the CRISPR/Cas system and use hands-on techniques to apply. On the first day of the course, we start with certain basic topics for better understanding. This is followed by illustrated explanations of the functionality, applications and specifics of the CRISPR/Cas system. We then draw the practical context from the example of a digital experiment, where the planning and selection of experimental components is explained and illustrated step by step. On the second day, the practical part follows with the application of techniques for manipulating genomic DNA in zygotes/embryos by microinjection and electroporation. Here, the participants are divided into two groups over two days.

Organizer: TransMed in cooperation with the Translational Animal Research Center (TARC)

Topics:  

Theoretical part:

• Introduction and basic
  • Tools of genetic engineering. Endonucleases in general, zinc finger nucleases, TALEN
  • Homologous recombination
  • DNA repair mechanisms specifically non-homologous end joining (NHEJ) and homology-directed repair (HDR)
• CRISPR/Cas
  • How CRISPR/Cas works and its history
  • Understanding CRISPR/Cas as a tool and its variants (Cas9, Cas12a, Cas13)
  • CRISPR/Cas applications
  • Advantages and disadvantages
• Digital experiment
  • Generation of a point mutation using CRISPR/Cas system in mouse zygotes
• Early embryogenesis (in preparation for the practical part)

Practical part:

• Thawing and handling of zygotes
• Microinjection - (pro)nucleus, cytoplasm into zygotes/embryos
• Electroporation of zygotes/embryos

Target Audience: TransMed's doctoral candidates, clinician scientists, and postdocs

Maximum number of participants: 12

Next Workshop: TBA

Venue: Seminar room at the TARC, Hans-Dieter-Hüsch-Weg 19, JGU Mainz

For the workshop,  1 TransMed CP for scientific skills training can be credited.