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ORNL researchers have engineered an innovative cloning system for flexible DNA assembly from universal libraries to generate multi-gene constructs (DNA synthesis). This cloning system adopts a pre-defined three nucleotides (TNT) signature and a buffer system for a quick one-pot reaction. The all-inclusive and ultra-flexible TNT-cloning system assembles functional constructs from a universal library that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, sequence homology, amplification or mutation of DNA fragments in order to work properly. The enhanced buffer formulation provides the capability for testing, and sharing, complex multi-gene circuitry composed from a variety of DNA fragments.
The system has also been engineered to shuttle between different biological systems. The easily transferable platform can be used to assemble the gene construct from a randomized library in vectors ready for transformation in a variety of hosts including bacteria, yeast, plant, and mammalian cells.
This technology offers gene assembly the power of harnessing large randomized libraries while also allowing for specific DNA building blocks to be catalogued for ease of access in the future or among research communities. In addition, the TNT-cloning system was specifically designed with the following advantages:
• Handles Large DNA Constructs- compatible with large nucleotide cassettes >50 kb in length (12kb fragments tested, ORNL).
• Highly Randomized and Populated Libraries- engineered to quickly assemble and select from large randomized libraries with reduced selection rounds.
• Gene Assembly Speed- the design streamlines steps for a system that is up to 80% faster than other assembly systems. Efficiency in steps and cloning speed increases with the number of assemblies. Use of high throughput and robotics has been validated with no drop in efficiency.
This technology was originally developed using federal funds and selected for further development under the laboratory's Technology Innovation Program (TIP). TIP supports technology development using royalties from existing technology licenses to accelerate the market readiness of high potential technologies.
License applications will be evaluated based on prospective partners' ability and commitment to successfully commercialize the technology. If a license is executed, the ORNL researchers developing the technology will be eligible to compete for additional TIP funding to further advance the technology.
Please contact Jennifer Caldwell, Group Leader, Technology Commercialization, Oak Ridge National Laboratory, 865.574.4180, [email protected], with any questions and to obtain a license application for this opportunity.
Intellectual Property
UTB ID 201403356, TNT-Cloning System, U.S. Patent Application 14/789,112 filed on July 1, 2015.
Attachment below includes the ORNL website (www.ornl.gov/partnerships/tnt-cloning-system) where additional information about this licensing opportunity, including technology summaries and patent information, is found.