Journal
SCIENCE
Volume 353, Issue 6304, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aag0511
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Funding
- National Institutes of Health [DP2 OD008435, P50 GM098792]
- Office of Naval Research
- National Science Foundation [MCB-1350625]
- Defense Advanced Research Projects Agency
- Center for Microbiome Informatics and Therapeutics
- NSF Expeditions in Computing Program [1522074]
- Natural Sciences and Engineering Research Council of Canada
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1350625] Funding Source: National Science Foundation
- Direct For Computer & Info Scie & Enginr
- Division of Computing and Communication Foundations [1522074] Funding Source: National Science Foundation
- Division of Computing and Communication Foundations
- Direct For Computer & Info Scie & Enginr [1521925] Funding Source: National Science Foundation
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The ability to record molecular events in vivo would enable monitoring of signaling dynamics within cellular niches and critical factors that orchestrate cellular behavior. We present a self-contained analog memory device for longitudinal recording of molecular stimuli into DNA mutations in human cells. This device consists of a self-targeting guide RNA (stgRNA) that repeatedly directs Streptococcus pyogenes Cas9 nuclease activity toward the DNA that encodes the stgRNA, enabling localized, continuous DNA mutagenesis as a function of stgRNA expression. We demonstrate programmable and multiplexed memory storage in human cells triggered by exogenous inducers or inflammation, both in vitro and in vivo. This tool, Mammalian Synthetic Cellular Recorder Integrating Biological Events (mSCRIBE), provides a distinct strategy for investigating cell biology in vivo and enables continuous evolution of targeted DNA sequences.
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