Frankfurt CRISPR/Cas Screening Center (FCSC)
The recently introduced CRISPR/Cas technology initiated an unprecedented biological revolution. As such, there is an increasing demand on gene editing technologies, including single and multiple gene knockouts, transcriptional activation/repression, targeted single nucleotide modification or the precise replacement of genetic loci for the purpose of tagging or modification.
To meet the increasing demand within the Institute and the local community, the IBC2 has built up the Frankfurt CRISPR/Cas Screening Center (FCSC). On the basis of collaboration, the platform supports both internal and external researchers with expertise, reagents and strategic advice on gene editing. The FCSC guides researches along their individual needs, ranging from single to multiple gene editing events, specific gene replacements, to the generation of custom-made gRNA libraries.
To generate high-quality gene editing reagents with unprecedented coverage, the FCSC mainly relies on the proprietary 3Cs technology, which was invented at the IBC2. The FCSC also contributes to further development of the technology for new applications.
The FCSC is closely connected with several research networks, including the CRC 1177 on “Molecular and Functional Characterization of Selective Autophagy”, the Excellence Cluster “Cardiopulmonary Institute (CPI)”, Frankfurt Cancer Institute (FCI) and Innovative Medicine Initiative (IMI).
Any requests for collaborations can be addressed to Koraljka Husnjak (+49 69 6301 5820).
The Frankfurt CRISPR/Cas Screening Platform (FCSP) is headed jointly by Ivan Dikic and Manuel Kaulich. In December 2015, Manuel was recruited to the IBC2 to build up an independent research group on cell cycle regulation. His technological expertise is in novel genome editing approaches, which he applies to address research questions related to cancer drug resistance and synthetic lethality. Following the enormous demand for CRISPR/Cas genome editing libraries within the local community, IBC2 Director Ivan took a strategic decision in December 2017 to set up a core facility serving the needs within and outside the Institute. Together, Ivan and Manuel are now leading a team of three scientists and one technical assistant to generate reagents and advising collaborators on the design and appropriate read-out of respective experiments.
Alkmini graduated from the University of Thessaly in Greece having a Bachelor’s degree in Biochemistry & Biotechnology, a Master’s degree in Molecular Applications of Molecular Medicine and a PhD degree in Biochemistry. During her PhD she investigated the regulation of the hypoxia-inducible factor by phosphorylation. Then she moved to Strasbourg for her postdoctoral research and worked at the Institute of Genetics and Molecular and Cellular Biology (IGBMC) with a focus on the role of chromatin and nuclear architecture in DNA repair. Alkmini joined FCSC in June 2019 as a postdoctoral research scientist.
Michaela Braun did her technical assistant (MTA) training in Heidelberg and works at the IBC2 since 2012 to support our scientific staff in the implementation and execution of their research projects. From January 2018 on, Michy has taken the challenge to join the team building up the FCSC.
Yves received his diploma in Biology and Computer Science from the Goethe University of Frankfurt. He developed an inducible shRNA system and an shRNA prediction algorithm for RNA interference (RNAi) to uncover the functions of oncogenes. Applying the inducible RNAi system, a transgenic animal model was implemented to demonstrate cancer addiction of Polo-like kinase 1. Working on the cell cycle at the Institute of Gynecology and Obstetrics in Frankfurt he used RNAi screening technology and high-throughput data analysis for improving drug target opportunities for ovarian cancer patients. In 2018 he joined the IBC2 as postdoctoral scientist building up the FCSC platform. Recently he designed an inducible and multiplex-based CRISPR/Cas9 system for conditional gene knockouts.
Unbiased and tailored CRISPR/Cas gRNA libraries by synthesizing covalently-closed-circular (3Cs) DNA. Wegner M, Husnjak K, Kaulich M. Bio-protocol (2020) 10(1): e3472. DOI: 10.21769/BioProtoc.3472
Circular synthesized CRISPR/Cas gRNAs for functional interrogations in the coding and noncoding genome. Wegner M, Diehl V, Bittl V, de Bruyn R, Wiechmann S, Matthess Y, Hebel M, Hayes MG, Schaubeck S, Benner C, Heinz S, Bremm A, Dikic I, Ernst A, Kaulich M. Elife ( 2019) 8. pii: e42549. doi: 10.7554/eLife.42549.
Wegner M, Diehl V, Bittl V, de Bruyn R, Wiechmann S, Matthess Y, Hebel M, Hayes MG, Schaubeck S, Benner C, Heinz S, Bremm A, Dikic I, Ernst A, Kaulich M. Circular synthesized CRISPR/Cas gRNAs for functional interrogations in the coding and noncoding genome. Elife. 2019 Mar 6;8. pii: e42549. doi: 10.7554/eLife.42549.
Wegner MS, Schömel N, Gruber L, Örtel SB, Kjellberg MA, Mattjus P, Kurz J, Trautmann S, Peng B, Wegner M, Kaulich M, Ahrends R, Geisslinger G, Grösch S. UDP-glucose ceramide glucosyltransferase activates AKT, promoted proliferation, and doxorubicin resistance in breast cancer cells. Cell Mol Life Sci. doi: 10.1007/s00018-018-2799-7.
Le Guerroué F, Eck F, Jung J, Starzetz T, Mittelbronn M, Kaulich M, Behrends C. Autophagosomal Content Profiling Reveals an LC3C-Dependent Piecemeal Mitophagy Pathway. Mol. Cell 2017. 68 (4): 786-796.e6.
de Bruyn R, Diehl V, Wegner M, Kaulich M. Präklinische Anwendungen von CRISPR/Cas - CRISPR/Cas applications in preclinical research. Pharmakon. 2017 4:258-264.
Wesely J, Steiner M, Schnütgen F, Kaulich M, Rieger MA, Zörnig M. Delayed Mesoderm and Erythroid Differentiation of Murine Embryonic Stem Cells in the Absence of the Transcriptional Regulator FUBP1. Stem Cells Int. 2017; 2017:5762301.
van Wijk SJL, Fricke F, Herhaus L, Gupta J, Hötte K, Pampaloni F, Grumati P, Kaulich M, Sou YS, Komatsu M, Greten FR, Fulda S, Heilemann M, Dikic I. Linear ubiquitination of cytosolic Salmonella Typhimurium activates NF-κB and restricts bacterial proliferation. Nat Microbiol. 2017 May 8;2:17066.
Key publications on CRISPR technology development of Manuel Kaulich during his postdoctoral tenure at UC San Diego in the Dowdy laboratory:
Meitinger F, Anzola JV, Kaulich M, Richardson A, Stender JD, Benner C, Glass CK, Dowdy SF, Desai A, Shiau AK, Oegema K. 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration. J Cell Biol. 2016 Jul 18;214(2):155-66.
Kaulich M, Lee YJ, Lönn P, Springer AD, Meade BR, Dowdy SF. Efficient CRISPR-rAAV engineering of endogenous genes to study protein function by allele-specific RNAi. Nucleic Acids Res. 2015 Apr 20;43(7):e45.
Kaulich M, Dowdy SF. Combining CRISPR/Cas9 and rAAV Templates for Efficient Gene Editing. Nucleic Acid Ther. 2015 Dec;25(6):287-96.