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Generalizing mechanisms of secondary structure dynamics in biopolymers

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  • معلومة اضافية
    • Contributors:
      Universität Leipzig
    • الموضوع:
      2023
    • Collection:
      Universität Leipzig: Qucosa
    • نبذة مختصرة :
      Secondary structure dynamics of biopolymers play a vital role in many of the complex processes within a cell. However, due to the substantial number of atoms in the involved biopolymers along with the multitude of interactions that occur between the molecules, understanding these processes in detail is challenging and often involves computationally demanding simulations. In this thesis, the secondary structure dynamics of three different biopolymer systems were modeled using a single approach, which is based on intuitive principles that facilitate the interpretation. To this end, the kinetic behavior of each system was experimentally determined, and described by simplified reaction schemes, which were then connected to Markov chain models encompassing all principal secondary structural conformations. Firstly, we investigated the toehold-mediated strand displacement reaction, which is widely applied in nanotechnology to create DNA-based nano-devices and biochemical reaction networks. Our model correctly described the impact of base pair mismatches on the kinetics of these reactions, as measured by bulk fluorescence experiments. Additionally, it revealed that incumbent dissociation, base pair fraying, and internal loop formation are important processes during strand displacement. Furthermore, we established two dissipative elements to enhance temporal control over toehold-mediated strand displacement reactions. The first element allowed a reversible and repeatable incumbent strand release, whereas the second element provided the possibility to start the displacement reaction after a programmable temporal delay. Secondly, we studied the target recognition by the CRISPR-Cas effector complex Cascade, a highly promising protein for applications in genome engineering. Our model successfully reproduced all aspects of the torque- and mismatch-dependent R-loop formation time by Cascade obtained by single-molecule torque and bulk fluorescence measurements. Furthermore, we demonstrated that the seed effect observed for ...
    • Relation:
      urn:nbn:de:bsz:15-qucosa2-901347; https://ul.qucosa.de/id/qucosa%3A90134; https://ul.qucosa.de/api/qucosa%3A90134/attachment/ATT-0/
    • الدخول الالكتروني :
      https://nbn-resolving.org/urn:nbn:de:bsz:15-qucosa2-901347
      https://ul.qucosa.de/id/qucosa%3A90134
      https://ul.qucosa.de/api/qucosa%3A90134/attachment/ATT-0/
    • Rights:
      info:eu-repo/semantics/openAccess
    • الرقم المعرف:
      edsbas.3553A7B9