|
|||||
|
|
Catalytic DNA-Assisted Mass Production of Arbitrary Single-Stranded DNA |
|
| Authors: | Qiao Zhang Dr. Kai Xia Meng Jiang Dr. Qingting Li Prof. Weigang Chen Prof. Mingzhe Han Wei Li Prof. Rongqin Ke Prof. Fei Wang Prof. Yongxing Zhao Prof. Yuehua Liu Prof. Chunhai Fan Prof. Hongzhou Gu |
| Affiliation: | 1. Fudan University Shanghai Cancer Center, and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Shanghai Stomatological Hospital, Fudan University, Shanghai, 200433 China These authors contributed equally to this work.;2. Fudan University Shanghai Cancer Center, and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Shanghai Stomatological Hospital, Fudan University, Shanghai, 200433 China Department of Chemical Biology, School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules, National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai, 201108 China Shanghai Frontier Innovation Research Institute, Shanghai, 201108 China These authors contributed equally to this work.;3. School of Medicine and School of Biomedical Science, Huaqiao University, Fujian, 362021 China These authors contributed equally to this work.;4. Fudan University Shanghai Cancer Center, and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Shanghai Stomatological Hospital, Fudan University, Shanghai, 200433 China Department of Chemical Biology, School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules, National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai, 201108 China;5. Frontier Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin, 300072 China;6. Fudan University Shanghai Cancer Center, and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Shanghai Stomatological Hospital, Fudan University, Shanghai, 200433 China;7. School of Medicine and School of Biomedical Science, Huaqiao University, Fujian, 362021 China;8. Department of Chemical Biology, School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules, National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai, 201108 China;9. Department of Pharmaceutics, School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and Diagnosis for Critical Diseases, and Key Laboratory of Advanced Drug Preparation Technologies, Zhengzhou University, Henan, 450001 China |
| Abstract: | Synthetic single-stranded (ss) DNA is a cornerstone for life and materials science, yet the purity, quantity, length, and customizability of synthetic DNA are still limiting in various applications. Here, we present PECAN, p aired-e nd c utting a ssisted by DN Azymes (DNA enzymes or deoxyribozymes), which enables mass production of ssDNA of arbitrary sequence (up to 7000 nucleotides, or nt) with single-base precision. At the core of PECAN technique are two newly identified classes of DNAzymes, each robustly self-hydrolyzing with minimal sequence requirement up- or down-stream of its cleavage site. Flanking the target ssDNA with a pair of such DNAzymes generates a precursor ssDNA amplifiable by pseudogene-recombinant bacteriophage, which subsequently releases the target ssDNA in large quantities after efficient auto-processing. PECAN produces ssDNA of virtually any terminal bases and compositions with >98.5 % purity at the milligram-to-gram scale. We demonstrate the feasibility of using PECAN ssDNA for RNA in situ detection, homology-directed genome editing, and DNA-based data storage. |
| Keywords: | DNA Synthesis DNAzyme Data Storage Genome Editing Padlock Probing |