Targeted DNA degradation using a CRISPR device stably carried in the host genome
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Caliando-2015-Targeted DNA.pdf
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652.88 KB
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Adobe PDF
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Author(s)
Caliando, Brian J.
Voigt, Christopher A.
Date Issued
May 2015
Journal
Nature Communications
Publisher
Nature Publishing Group
Citation
Caliando, Brian J., and Christopher A. Voigt. “Targeted DNA Degradation Using a CRISPR Device Stably Carried in the Host Genome.” Nature Communications 6 (May 19, 2015): 6989. © 2015 Macmillan Publishers Limited
Version
Final published version
Abstract
Once an engineered organism completes its task, it is useful to degrade the associated DNA to reduce environmental release and protect intellectual property. Here we present a genetically encoded device (DNAi) that responds to a transcriptional input and degrades user-defined DNA. This enables engineered regions to be obscured when the cell enters a new environment. DNAi is based on type-IE CRISPR biochemistry and a synthetic CRISPR array defines the DNA target(s). When the input is on, plasmid DNA is degraded 10[superscript 8]-fold. When the genome is targeted, this causes cell death, reducing viable cells by a factor of 10[superscript 8]. Further, the CRISPR nuclease can direct degradation to specific genomic regions (for example, engineered or inserted DNA), which could be used to complicate recovery and sequencing efforts. DNAi can be stably carried in an engineered organism, with no impact on cell growth, plasmid stability or DNAi inducibility even after passaging for >2 months.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Synthetic Biology Center
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1038/ncomms7989
