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Stability and Activity of the 10-23 DNAzyme Under Molecular Crowding Conditions.
Kirchgässler, Nina; Rosenbach, Hannah; Span, Ingrid.
Affiliation
  • Kirchgässler N; Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany.
  • Rosenbach H; Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany.
  • Span I; Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany. ingrid.span@hhu.de.
Methods Mol Biol ; 2439: 79-89, 2022.
Article in En | MEDLINE | ID: mdl-35226316
DNAzymes are biocatalysts that have been selected in vitro and their function inside cells (in vivo) is extremely low. Thus, almost all studies have been carried out in diluted solutions (in vitro). The cellular presence of molecules such as amino acids, polypeptides, alcohols, and sugars introduces forces that modify the kinetics and thermodynamics of DNAzyme-mediated catalysis. The crowded intracellular environment referred to as molecular crowding can be mimicked by adding high concentrations of natural or synthetic macromolecules to the reaction conditions. Here, we investigate the activity of the 10-23 DNAzyme and the stability of the DNAzyme:RNA complex under molecular crowding conditions. Therefore, we use a Förster resonance energy transfer (FRET)-based activity assay in combination with denaturing urea polyacrylamide gel electrophoresis and circular dichroism (CD) spectroscopy.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: DNA, Catalytic Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2022 Document type: Article Affiliation country: Germany Country of publication: United States

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: DNA, Catalytic Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2022 Document type: Article Affiliation country: Germany Country of publication: United States