biostudies-literatureZhang CNational Natural Science Foundation of ChinaNational Key Research and Development Program of China6364-6374https://www.ebi.ac.uk/biostudies/studies/S-EPMC8216280biostudies-literatureUnknown49(11)DNA-hydrolyzing DNAs represent an attractive type of DNA-processing catalysts distinctive from the protein-based restriction enzymes. The innate DNA property has enabled them to readily join DNA-based manipulations to promote the development of DNA biotechnology. A major in vitro selection strategy to identify these DNA catalysts relies tightly on the isolation of linear DNAs processed from a circular single-stranded (ss) DNA sequence library by self-hydrolysis. Herein, we report that by programming a terminal hybridization stem in the library, other than the previously reported classes (I & II) of deoxyribozymes, two new classes (III & IV) were identified with the old selection strategy to site-specifically hydrolyze DNA in the presence of Zn2+. Their representatives own a catalytic core Nucleic acids researchNew DNA-hydrolyzing DNAs isolated from an ssDNA library carrying a terminal hybridization stem.PMC82162802167305091859104818611380042020YFA0908901Li WHe YGu HXu TLi QZhang CfalseNew DNA-hydrolyzing DNAs isolated from an ssDNA library carrying a terminal hybridization stem.DNA-hydrolyzing DNAs represent an attractive type of DNA-processing catalysts distinctive from the protein-based restriction enzymes. The innate DNA property has enabled them to readily join DNA-based manipulations to promote the development of DNA biotechnology. A major in vitro selection strategy to identify these DNA catalysts relies tightly on the isolation of linear DNAs processed from a circular single-stranded (ss) DNA sequence library by self-hydrolysis. Herein, we report that by programming a terminal hybridization stem in the library, other than the previously reported classes (I & II) of deoxyribozymes, two new classes (III & IV) were identified with the old selection strategy to site-specifically hydrolyze DNA in the presence of Zn2+. Their representatives own a catalytic core 2021-01-01T00:00:00Z2021 Jun2025-04-04T20:18:19.464Z2022-02-10T15:47:17.448ZS-EPMC82162803405747610.1093/nar/gkab439