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Properties of an LNA-modified ricin RNA aptamer
Authors:Förster Charlotte  Zydek Martin  Rothkegel Maika  Wu Zhiyang  Gallin Claudia  Geßner Reinhard  Lisdat Fred  Fürste Jens P
Institution:1. Institute for Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany;2. Clinic for Oral and Maxillofacial Surgery, Henriettenstiftung, Hannover, Germany;3. Clinic for General- and Visceral Surgery, Section of Pediatric Surgery, University Clinic Ulm, Ulm, Germany;4. Department of Trauma Surgery, Hannover Medical School, Hannover, Germany;5. Department of Anatomy and Cell Biology, Martin Luther University, Faculty of Medicine, Halle (Saale), Germany
Abstract:'Locked nucleic acids' (LNAs) are sugar modified nucleic acids containing the 2'-O-4'C-methylene-β-D-ribofuranoses. The substitution of RNAs with LNAs leads to an enhanced thermostability. Aptamers are nucleic acids, which are selected for specific target binding from a large library pool by the 'SELEX' method. Introduction of modified nucleic acids into aptamers can improve their stability. The stem region of a ricin A chain RNA aptamer was substituted by locked nucleic acids. Different constructs of the LNA-substituted aptamers were examined for their thermostability, binding activity, folding and RNase sensitivity as compared to the natural RNA counterpart. The LNA-modified aptamers were active in target binding, while the loop regions and the adjacent stem nucleotides remained unsubstituted. The thermostability and RNase resistance of LNA substituted aptamers were enhanced as compared to the native RNA aptamer. This study supports the approach to substitute the aptamer stem region by LNAs and to leave the loop region unmodified, which is responsible for ligand binding. Thus, LNAs possess an encouraging potential for the development of new stabilized nucleic acids and will promote future diagnostic and therapeutic applications.
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