Titel der Forschungsarbeit: Genetic invalidation of the SARS-CoV-2-receptor ACE2 in a colon cancer cell line by CRISPR/Cas9 mutagenesis
Fakultät: Fakultät für Medizin
Lehrstuhl: Klinik und Poliklinik für Chirurgie
Betreuung: Prof. Dr. rer. nat. Klaus-Peter Janssen, PD Dr. Jeannine Bachmann
Abstract der Forschungsarbeit
Angiotensin I converting enzyme 2 (ACE2) is a protein expressed in tissue in several parts of the human body ranging from the intestinal tract to the bronchus. As a regulator of blood pressure and intestinal homeostasis, it plays an important role in different biochemical systems in the human body. Moreover, it has recently gained interest as a cellular receptor for the novel SARS-CoV-2 virus that is causing a global pandemic. Although the research regarding the virus is primarily focused on the development of a vaccine, studying the way the virus enters cells is of high relevance as well. This, as well as the precise mechanisms and function of ACE2 in intestinal epithelial cells, has yet to be fully researched.
Therefore, this research project was focused on creating a cell line that allows further research into the key roles of ACE2. This was achieved by a genetic invalidation of the protein in the human colon cancer cell line HCT116 using the CRISPR/Cas9 technique. After designing a sgRNA and cloning a vector carrying the genes causing the invalidation of ACE2 in the cells, HCT116 cells were transfected with this vector. By deactivating ACE2 in the human colon cancer cells, the consequences of malfunctioning or a lack of the protein can be investigated and consequently, deductions regarding the complex role of ACE2 can be made. This is, for example, very important for the development of treatments of diseases interacting with the protein as for instance COVID-19 with the SARS-CoV-2 pathogen.