Engineered Genetic Device for the Controlled Destruction of Genomic and Plasmid DNA

Technology #15586

Questions about this technology? Ask a Technology Manager

Download Printable PDF

Image Gallery
DNA destruction device (DDD) in the context of an E. coli cell. Experimental design assesses the DDD degrading an existing target DNA.  A bar graph presenting a time-course for the loss of the ampicillin resistance-conferring plasmid.
Categories
Inventors
Professor Christopher Voigt
Department of Biological Engineering, MIT
External Link (mit.edu)
Brian Caliando
Department of Biological Engineering, MIT
Managed By
Jon Gilbert
MIT Technology Licensing Officer
Patent Protection

Genetic device for the controlled destruction of dna

PCT Patent Application WO 2014-071235

Genetic device for the controlled destruction of dna

US Patent Pending 2015-0315576

Applications

This invention enables target specific DNA destruction in a highly controlled and regulated manner.

Problem Addressed

Current methods for removing genetic material from a living prokaryotic host incompletely removes the genetic element from the population at large. Furthermore, targeting specificities of all other known prokaryotic nucleases capable of degrading DNA in vivo are encoded in their protein sequences, making it difficult if not impossible to readily alter the intended target sequence specificity without extensive mutagenesis and screening. This invention is a DNA destruction device (DDD) that uses CRISPR biology to address both of these issues; it prevents propagation of a DNA sequence to progeny while actively degrading these elements, lowering the total copy number of existing elements in the population gene pool and can be simply and quickly re-targeted to destroy specific DNA.

Technology

The device is composed of two essential components engineered from E. coli genes. The first is the CRISPR array, a synthetic DNA sequence containing series of unique 32-bp fragments interspersed between identical 29-bp palindromic repeats of specific sequence. The array's spacers encode the device's targeting information. The second component is a cas gene (cas3 or casABCDE), which encodes the device's actuator functions, specifically the enzymatic machinery sufficient for catalyzing DNA degradation. Both of these essential components are put under the artificial control of independent inducible promoters, which allow for user controlled activation of the device in response to specific combination of environmental or chemical signals. The device is highly effective at removing specific genetic elements from cells when and where desired.  Some examples of targeted genetic elements could be exogenous dangerous or confidential DNA sequences or DNA being used by the host cell for protein or other production.

Advantages

  • Allows for the user-controlled, irreversible removal of virtually any genetic element