Freiburg iGEM 2009
1. Short Summary Project
Universal Endonuclease – Cutting Edge Technology
Gene technology is driven by the use of restriction endonucleases. Yet, constraints of limited sequence length and variation recognized by available restriction enzymes pose a major roadblock for synthetic biology. We developed the basis for universal restriction enzymes, primarily for routine cloning but also with potential for in vivo applications. We use a nucleotide cleavage domain fused to a binding domain, which recognizes a programmable adapter that mediates binding to DNA and thus cleavage. As adapter we use readily available modified oligonucleotides, as binding domain anticalins and as cleavage domain FokI moieties engineered for heterodimerization and activity. For cloning, this universal enzyme has merely to be mixed with the sequence specific oligonucleotide and the target DNA. Binding and release are addressed with thermocycling. Additionally, we provide concepts for in vivo applications by external adapter delivery, activity regulation by photo-switching, as well as for modifying an argonaute protein towards a DNA endonuclease.
2. Team:
Advisors: Dr. Katja Arndt, Dr. Kristian Müller, Sven Hagen, Janina Speck , Tobias Baumann
Students: Anika Lückoff, Carolin Nieder, Christoph Deeg, Gerrit Gresch, Hannes Breddermann, Isabel Lohrmann, Julia Rennertz, Laura Eichstädter, Manuel Hiss, Maximilian Müller, Sarah Bambach, Timo Lorenz, Rüdiger Trojok, Sascha Duttke
3. Achievements:
Freiburg bioware team: finalist, best BioBrick engineered, best poster, gold medal
Freiburg software team: first German software team, silver medal