Electro-addressable in vitro transcription system for random access and rewriting in dynamic DNA storage

Due to the rapid development of parallel array-based DNA synthesis, DNA storage has gradually demonstrated the potential to replace silicon-based storage. However, current methods typically require cutting the array DNA after synthesis for subsequent operations (such as preservation in small tubes or fixation on a chip), which prevents direct reading and hinders its practical application. This study developed a universal random access method based on electrode array chips, which can not only achieve in-situ random access without additional processing steps after DNA array chemical synthesis, but also meet the requirements of random access after array synthesis and sequence cutting fixed on the chip. By simulating the direction requirement of DNA synthesis from 3' to 5', the DNA is only desalted after chemical synthesis, and the 3' end of the DNA is chemically bonded to the surface of the electrode chip. Subsequently, physical addressing is achieved through rapid hybridization with the electro-assisted T7 promoter, and information replication is carried out through in vitro transcription. This method effectively reduces the complex computations and low base usage efficiency caused by traditional primer sequence addressing through electro-addressing and the need for only a pair of primers; meanwhile, the replication process under isothermal conditions meets the demand for one-time writing and multiple readings; in addition, the synergistic effect of exonuclease and DNA ligase also supports multiple erasures and rewrites of information. In conclusion, compared with previous DNA-based data storage methods, this method demonstrates superior compatibility and stability, providing a new technical path for the future realization of miniaturized and automated DNA data storage devices.