Gene knock-out means that the function of a specific gene, and consequently that of its encoded protein, is disrupted by a specific mutation. Gene knock-in means that the specific target gene and its protein gain a new function because of an engineered mutation. By using the resulting CRISPR/Cas cell clones, the function of specific genes and proteins can be studied in detail within the cellular context.
After its publication in 2012, the CRISPR/Cas gene editing system revolutionized molecular biology within only a few years (CRISPR = clustered regularly interspaced short palindromic repeats; see also our Introduction page). The increase in publications using CRISPR/Cas is only comparable to what was seen in the eighties following the development of the polymerase chain reaction (PCR) or in the nineties following the introduction of the green fluorescent protein (GFP). The Cell Engineering Facility of the FMP will make gene knock-outs and knock-ins in various target cells using state-of-the art CRISPR/Cas techniques. Other applications may be possible following consultation. Cell clones will be characterized by DNA sequencing and the genotype will be analyzed to confirm modification of both alleles. Western blotting will be done in case you could provide a specific antibody.
An overview of the methodology we use can be found on the “Methods” page. If you are already familiar with CRISPR/Cas, you may immediately proceed to the “Order” page. Depending on the amount of simultaneous orders, clones should be ready within 8-12 weeks depending on cell type. If you use the system for the first time or need an overview of its history, function and applications, you may visit our “Introduction” page first. If you have any questions, please get in touch.
In agreement with the FMP directors, we will charge will charge 1,000 Euro € for a CRISPR/Cas cell clone (knock-in or knock out) when the clone is confirmed.
In addition to CRISPR/Cas gene editing, the Cell Engineering Facility is also able to make stable transfected cell clones using various cell lines upon request.
Ralf Schülein studied biology in Würzburg and did his doctorate there at the Chair of Microbiology. As a post-doc, he worked for three years at the Rudolf Buchheim Institute for Pharmacology at the University of Giessen. He has headed a research group at the FMP since 1997 and the Cell Engineering Facility since 2020. The habilitation in pharmacology and toxicology took place at the former Faculty of Medicine at the Freie Universität Berlin. In 2014, Ralf Schülein was appointed adjunct professor at the Charite University Medicine Berlin. He teaches medical students in pharmacology and toxicology. He also has a teaching position at the Freie Universität Berlin.