Molecular recordings by directed CRISPR spacer acquisition

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 intersp...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2016-07, Vol.353 (6298), p.463-463
Main Authors: Shipman, Seth L., Nivala, Jeff, Macklis, Jeffrey D., Church, George M.
Format: Article
Language:English
Subjects:
Acquisitions
Bacteria
Clustered Regularly Interspaced Short Palindromic Repeats
CRISPR-Associated Proteins - genetics
CRISPR-Associated Proteins - metabolism
CRISPR-Cas Systems
Deoxyribonucleic acid
Devices
Directed Molecular Evolution
Endodeoxyribonucleases - genetics
Endodeoxyribonucleases - metabolism
Endonucleases - genetics
Endonucleases - metabolism
Escherichia coli Proteins - genetics
Escherichia coli Proteins - metabolism
Evolution
Genome, Bacterial
Genomics
Literary Devices
Marketing
Molecular biology
Recording
RESEARCH ARTICLE SUMMARY
Spacers
Synthetic Biology - methods
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Summary: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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.aaf1175