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- Title
Zn2+-dependent DNAzymes that cleave all combinations of ribonucleotides.
- Authors
Inomata, Rika; Zhao, Jing; Miyagishi, Makoto
- Abstract
Although several DNAzymes are known, their utility is limited by a narrow range of substrate specificity. Here, we report the isolation of two zinc-dependent DNAzymes, ZincDz1 and ZincDz2, which exhibit compact catalytic core sequences with highly versatile hydrolysis activity. They were selected through in vitro selection followed by deep sequencing analysis. Despite their sequence similarity, each DNAzyme showed different Zn2+-concentration and pH-dependent reaction profiles, and cleaved the target RNA sequences at different sites. Using various substrate RNA sequences, we found that the cleavage sequence specificity of ZincDz2 and its highly active mutant ZincDz2-v2 to be 5′-rN↓rNrPu-3′. Furthermore, we demonstrated that the designed ZincDz2 could cut microRNA miR-155 at three different sites. These DNAzymes could be useful in a broad range of applications in the fields of medicine and biotechnology. Inomata et al. isolate zinc-dependent DNAzymes with compact catalytic core sequences displaying highly versatile hydrolysis activity. The cleavable substrate variation for the DNAzymes (ZincDz2 and ZincDz2-v2) is very broad and the authors further demonstrate their utility by showing ZincDz2 can hydrolyze miR-155 in vitro at three different sites. With their broad substrate tolerance, this study advances the DNAzyme field for RNA cleavage applications.
- Subjects
DEOXYRIBOZYMES; RIBONUCLEOTIDES; BIOCHEMICAL substrates; NUCLEOTIDE sequence; HYDROLYSIS
- Publication
Communications Biology, 2021, Vol 4, Issue 1, p1
- ISSN
2399-3642
- Publication type
Article
- DOI
10.1038/s42003-021-01738-6