Thermo‐Responsive Living Microspheroids Enable a Regenerative Living Disk–Drive System for DNA Data Storage

Abstract

ABSTRACT DNA offers exceptional information density and long‐term stability, yet its practical deployment is limited by destructive readout and the absence of a reusable, physically addressable architecture that connects nanoscale molecular information with macroscale device‐level data organization. Here, we present a regenerative Living Disk–Drive system based on thermo‐responsive engineered living memory microspheroids (ELMMs), in which data‐encoded bacteria are encapsulated as discrete, file‐level living storage units. Each ELMM contains a clonal bacterial population carrying both an information plasmid, which encodes 26 × 26 pixel icon payloads and one‐ to three‐color intracellular fluorescent retrieval indices, and a help plasmid that enables CRISPR–Cas12a/λ‐Red rewriting of the data sequence and retrieval tag. A lyophilized ELMM database forms the Living Disk, which is coupled to an Optical Retriever and desktop‐scale Living Drive for closed‐loop retrieval, regeneration, and database replenishment. Released bacteria regrow for downstream readout or rewriting, while a fraction is re‐encapsulated into new ELMMs. The tested system retains retrieval, regrowth, and sequence recovery after four months of ambient dry storage and 13 lyophilization–rehydration cycles. Model‐based performance estimates are reported only as theoretical architecture‐level bounds. These results establish an experimentally bounded yet extensible architecture for physically manageable and regenerative DNA memory.