Journal
SCIENCE
Volume 353, Issue 6298, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aaf1175
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Funding
- National Institute on Aging [5T32AG000222]
- National Institute of Mental Health [5R01MH103910]
- National Human Genome Research Institute [5P50HG005550]
- National Institute of Neurological Disorders and Stroke [5R01NS045523]
- Allen Distinguished Investigator Award from the Paul G. Allen Family Foundation
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The ability to write a stable record of identified molecular events into a specific genomic locus would enable the examination of long cellular histories and have many applications, ranging from developmental biology to synthetic devices. We show that the type I-E CRISPR ( clustered regularly interspaced short palindromic repeats)-Cas system of Escherichia coli can mediate acquisition of defined pieces of synthetic DNA. We harnessed this feature to generate records of specific DNA sequences into a population of bacterial genomes. We then applied directed evolution so as to alter the recognition of a protospacer adjacent motif by the Cas1-Cas2 complex, which enabled recording in two modes simultaneously. We used this system to reveal aspects of spacer acquisition, fundamental to the CRISPR-Cas adaptation process. These results lay the foundations of a multimodal intracellular recording device.
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