Course Identification

Adapting CRISPR-Cas9-derived tools for site-specific genome engineering

Lecturers and Teaching Assistants

Dr. Dan Michael
Dr. Stav Kozlovski, Dr. Neta Degani, Dr. Noa Furth

Course Schedule and Location

Second Semester
6/5-7/5- FGS room B, 12/5-16/5, 19/5-22/5, 26/5; 13:00-18:00, FGS, Lab

Field of Study, Course Type and Credit Points

Life Sciences: Laboratory; Elective; 1.00 points
Life Sciences (Molecular and Cellular Neuroscience Track): Laboratory; Elective; 1.00 points
Life Sciences (Brain Sciences: Systems, Computational and Cognitive Neuroscience Track): Laboratory; Elective; 1.00 points
Life Sciences (Computational and Systems Biology Track): Laboratory; Elective; 1.00 points


Personal project and lab report are expected to be accomplished during the course of the lab.





Language of Instruction


Registration by


Attendance and participation


Grade Type

Pass / Fail

Grade Breakdown (in %)

Personal project and lab report

Evaluation Type


Scheduled date 1


Estimated Weekly Independent Workload (in hours)



Various CRISPR-Cas9-derived tools enable genome engineering, allowing efficient gene knock-out, knock-in, and site-specific regulation of epigenetic events. The course will provide hands-on training in genome editing using 3 generations of CRISPR-Cas9 tools. Thus, Cas9-expressing plasmids, Cas9 RNA, and the Cas9 protein will be employed, along with an appropriate single-guide RNA (sgRNA) to knock-out, knock-in, and edit at specific genomic loci. This course will emphasize training to design the necessary tools and to produce the sgRNA of interest and Cas9 RNA by in-vitro transcription as well as using ribo-nuclear protein (RNP) complexes. Transfection will be used side by side with electroporation to appreciate the importance of their optimization for efficient engineering outcomes. Assays for detecting gene modifications and clone validation will be employed.

To accompany the practical part, we will discuss various relevant topics, and to name just a few, we will study the emergence of several CRISPR-derived technologies as a vital discipline relevant for research and biotechnology in cells and organisms alike. This includes the current, yet limited, knowledge of the cellular DNA repair machineries that underlie efficient genomic modifications and the basic knowledge required for efficient gene knock-out and knock-in that should be characterized by minimal undesired “off-targets”.

Finally, participants will design their sgRNA of choice, produce it, and they will generate sufficient Cas9 RNA to allow them to initiate their own projects back at their mother laboratories. 

Learning Outcomes

Upon successful completion of the course, students will be able to:

  1. Appreciate the biochemical principles underlying the CRISPR-Cas9-derived technologies for genome engineering as well as the cellular machineries that are associated with genome editing.
  2. Demonstrate familiarity with designing and employing genome editing tools and methodologies optimized for their needs.
  3. Master knock out, knock-in and editing using 3 generations of Cas9 sources, as well as screening and validation of the engineered cells.
  4. Choose the right collection of gene editing tools and the right delivery mode for the cell type or organism of interest.

Reading List