Catalytic DNA‐Assisted Mass Production of Arbitrary Single‐Stranded DNA. (22nd December 2022)

• Record Type: Journal Article
• Authors: Zhang, Qiao; Xia, Kai; Jiang, Meng; Li, Qingting; Chen, Weigang; Han, Mingzhe; Li, Wei; Ke, Rongqin; Wang, Fei; Zhao, Yongxing; Liu, Yuehua; Fan, Chunhai; Gu, Hongzhou
• Is Part Of: Angewandte Chemie. Volume 135:Number 5(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-12-22
• Subjects: DNA Synthesis -- DNAzyme -- Data Storage -- Genome Editing -- Padlock Probing; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.202212011
• Languages: English
• ISSNs: 0044-8249

Abstract

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. Abstract : Based on two newly identified classes of DNAzymes, we developed p aired‐e nd c utting a ssisted by DN Azymes (PECAN) for mass production of scar‐free single‐stranded DNA (ssDNA). We demonstrated the feasibility of using PECAN ssDNA for in situ RNA detection, homology‐directed genome editing, and DNA‐based data storage, and highlighted the advantages of using high‐quality PECAN ssDNA in cutting‐edge biotechnology.