AbstractIn the face of exponential data growth, DNA‐based storage offers a promising solution for preserving big data. However, most existing DNA storage methods, akin to traditional block printing, require costly chemical synthesis for each individual data file, adopting a sequential, one‐time‐use synthesis approach. To overcome these limitations, a novel, cost‐effective “DNA‐movable‐type storage” system, inspired by movable type printing, is introduced. This system utilizes prefabricated DNA movable types‐short, double‐stranded DNA oligonucleotides encoding specific payload, address, and checksum data. These DNA‐MTs are enzymatically ligated/assembled into cohesive sequences, termed “DNA movable type blocks,” streamlining the assembly process with the automated BISHENG‐1 DNA‐MT inkjet printer. Using BISHENG‐1, 43.7 KB of data files are successfully printed, assembled, stored, and accurately retrieved in diverse formats (text, image, audio, and video) in vitro and in vivo, using only 350 DNA‐MTs. Notably, each DNA‐MT, synthesized once (2 OD), can be used up to 10000 times, reducing costs to $122/MB—outperforming existing DNA storage methods. This innovation circumvents the need to synthesize entire DNA sequences encoding files from scratch, offering significant cost and efficiency advantages. Furthermore, it has considerable untapped potential to advance a robust DNA storage system, better meeting the extensive data storage demands of the big‐data era.
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