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Molecular recordings by directed CRISPR spacer acquisition
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Molecular recordings by directed CRISPR spacer acquisition
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Journal Article
Molecular recordings by directed CRISPR spacer acquisition
2016
Overview
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.
Publisher
American Association for the Advancement of Science,The American Association for the Advancement of Science
Subject
/ Bacteria
/ Clustered Regularly Interspaced Short Palindromic Repeats
/ CRISPR-Associated Proteins - genetics
/ CRISPR-Associated Proteins - metabolism
/ Devices
/ Directed Molecular Evolution
/ DNA
/ E coli
/ Endodeoxyribonucleases - genetics
/ Endodeoxyribonucleases - metabolism
/ Escherichia coli Proteins - genetics
/ Escherichia coli Proteins - metabolism
/ Genomics
/ Mutants
/ Spacers
