New Deoxyribozymes for the Native Ligation of RNA

Molecules

Sandra Lange¹  Carolin P. M. Scheitl²  Claudia Höbartner² 

[1] Agricultural Center, BASF SE, Speyerer Str 2, 67117 Limburgerhof, Germany;Institute of Organic Chemistry, University of Würzburg, Am Hubland, 97074 Würzburg, Germany;

关键词: RNA ligation;    DNA catalysis;    in vitro selection;    deoxyribozyme;   

DOI  :  10.3390/molecules25163650

来源: DOAJ

【 摘 要 】

Deoxyribozymes (DNAzymes) are small, synthetic, single-stranded DNAs capable of catalyzing chemical reactions, including RNA ligation. Herein, we report a novel class of RNA ligase deoxyribozymes that utilize 5′-adenylated RNA (5′-AppRNA) as the donor substrate, mimicking the activated intermediates of protein-catalyzed RNA ligation. Four new DNAzymes were identified by in vitro selection from an N₄₀ random DNA library and were shown to catalyze the intermolecular linear RNA-RNA ligation via the formation of a native 3′-5′-phosphodiester linkage. The catalytic activity is distinct from previously described RNA-ligating deoxyribozymes. Kinetic analyses revealed the optimal incubation conditions for high ligation yields and demonstrated a broad RNA substrate scope. Together with the smooth synthetic accessibility of 5′-adenylated RNAs, the new DNA enzymes are promising tools for the protein-free synthesis of long RNAs, for example containing precious modified nucleotides or fluorescent labels for biochemical and biophysical investigations.

【 授权许可】

Unknown