High-throughput in vivo DNA data encoding by multiplexed conjugationand orthogonal base editing
Louise F. H. FunkeiD, Zhangyuan LiniD, Wenjun LiaoiD, Yutong JiiD, Chueh Loo PohiD
Abstract
Bacterial conjugation and CRISPR base editing offer complementary routes for programmable DNA transfer and heritable genomic writing, but their combined potential for information encoding in living cells remains unexplored. Here, we introduce CONCORD, a bacterial encoding platform that couples multiplexed bacterial conjugation to genomic base editing for synthesis-free in vivo DNA data writing. A recipient Writer strain expresses the base-editing machinery and is equipped with a writing template, whereas reusable donor Guide strains deliver the encoding sgRNA signals by conjugation. Using this setup, we encode binary information into genomic DNA in a high-throughput manner. Moreover, we systematically evaluated multiple orthogonal base editors for an expanded encoding repertoire and improved data recovery through redundancy encoding. We deploy the system to encode 45,689 bits into DNA including two culturally and historically significant artifacts: a greeting from NASA's Voyager Golden Record and a photograph of Navajo petroglyphs and recovered these datasets with 100% and 99.5% accuracy, respectively, without computational error correction. Connecting and engineering the unique mating and immune system of microbial cells we have established a framework for programmable information writing in bacterial populations, with applications for biological event recording and sustainable DNA data storage.
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