An RNA‐Cleaving DNAzyme That Requires an Organic Solvent to Function

Author:

Chang Tianjun1ORCID,Li Guangping1ORCID,Chang Dingran2ORCID,Amini Ryan2,Zhu Xiaoni1ORCID,Zhao Tongqian1,Gu Jimmy2,Li Zhongping3ORCID,Li Yingfu2ORCID

Affiliation:

1. School of Resources and Environment Henan Polytechnic University Jiaozuo 454000 China

2. Department of Biochemistry and Biomedical Sciences McMaster University Hamilton Ontario L8S 4K1 Canada

3. Institute of Environmental Science Shanxi University Taiyuan 030006 China

Abstract

AbstractEngineering functional nucleic acids that are active under unusual conditions will not only reveal their hidden abilities but also lay the groundwork for pursuing them for unique applications. Although many DNAzymes have been derived to catalyze diverse chemical reactions in aqueous solutions, no prior study has been set up to purposely derive DNAzymes that require an organic solvent to function. Herein, we utilized in vitro selection to isolate RNA‐cleaving DNAzymes from a random‐sequence DNA pool that were “compelled” to accept 35 % dimethyl sulfoxide (DMSO) as a cosolvent, via counter selection in a purely aqueous solution followed by positive selection in the same solution containing 35 % DMSO. This experiment led to the discovery of a new DNAzyme that requires 35 % DMSO for its catalytic activity and exhibits drastically reduced activity without DMSO. This DNAzyme also requires divalent metal ions for catalysis, and its activity is enhanced by monovalent ions. A minimized, more efficient DNAzyme was also derived. This work demonstrates that highly functional, organic solvent‐dependent DNAzymes can be isolated from random‐sequence DNA libraries via forced in vitro selection, thus expanding the capability and potential utility of catalytic DNA.

Funder

Natural Sciences and Engineering Research Council of Canada

Chinese Government Scholarship

National Natural Science Foundation of China

Publisher

Wiley

Subject

General Chemistry,Catalysis

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