| NSF Org: |
TI Translational Impacts |
| Recipient: |
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| Initial Amendment Date: | September 8, 2016 |
| Latest Amendment Date: | September 8, 2016 |
| Award Number: | 1640678 |
| Award Instrument: | Standard Grant |
| Program Manager: |
Jesus Soriano Molla
jsoriano@nsf.gov (703)292-7795 TI Translational Impacts TIP Directorate for Technology, Innovation, and Partnerships |
| Start Date: | September 15, 2016 |
| End Date: | February 28, 2018 (Estimated) |
| Total Intended Award Amount: | $200,000.00 |
| Total Awarded Amount to Date: | $200,000.00 |
| Funds Obligated to Date: |
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| History of Investigator: |
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| Recipient Sponsored Research Office: |
77 MASSACHUSETTS AVE CAMBRIDGE MA US 02139-4301 (617)253-1000 |
| Sponsor Congressional District: |
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| Primary Place of Performance: |
77 Massachusetts Ave Cambridge MA US 02139-4301 |
| Primary Place of
Performance Congressional District: |
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| Unique Entity Identifier (UEI): |
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| Parent UEI: |
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| NSF Program(s): | Accelerating Innovation Rsrch |
| Primary Program Source: |
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| Program Reference Code(s): |
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| Program Element Code(s): |
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| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.084 |
ABSTRACT
This PFI: AIR Technology Translation project focuses on translating next generation microfluidic genetic transformation technologies to fill the need for high throughput generation of genetically modified microorganisms. This system will allow metabolic engineers to more rapidly develop microorganisms for the production of various bioengineered chemicals and materials. This novel genetic transformation system is important because the fields of synthetic biology and genetic engineering are currently limited by the ability to re-program microorganisms with foreign DNA. This project will result in a prototype high throughput genetic transformation platform to demonstrate the utility of the system. This genetic transformation platform will be able to process microorganisms nearly one hundred times faster than the current state of the art.
This project addresses the following technology gap(s) as it translates from research discovery toward commercial application. Electroporation, cell permeabilization using pulsed electric fields, is an efficient way to deliver DNA constructs into microorganisms for genetic engineering and synthetic biology applications. Standard electroporation protocols involve parallel plate cuvettes to expose cell and DNA samples to uniform electric fields. However, testing different electroporation conditions involves hundreds of experiments (e.g. varying DNA construct, cell type, buffer composition), which is slow, labor-intensive, and expensive. This project aims to develop a prototype high throughput platform for genetic transformation of bacteria using microfluidic flow-through electroporation. The prototype device will be operated in 1) large scale production of bioengineered chemicals and 2) in a discovery mode to identify optimal transformation protocols for genetic engineering. This scalable technology will accelerate the development of new bioengineered chemicals that can be manufactured in a renewable manner.
Personnel involved in this project, research scientists, postdocs, and undergraduate students, will receive innovation, entrepreneurship, and technology translation experiences through participation in prototype design, customer interviews, business plan competitions, engaging with mentors, and establishing a company to commercialize the technology. The project engages the MIT Venture Mentoring Services and the MIT Innovation Initiative to guide commercialization aspects in this technology translation effort from research discovery toward commercial reality.
PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH
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PROJECT OUTCOMES REPORT
Disclaimer
This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.
We have developed a microfluidic electroporation technology that is scalable and has the potential for high-throughput genetic engineering of cells. This system will help bring high value products to market such as new biologically derived therapeutics and biomaterials. Electroporation is the most robust method of genetic modification but is primarily performed manually, leading to slow, unreliable, and low throughput genetic engineering. The Scalable Pipette Tip for High-Throughput Genetic Engineering we have developed in conjunction with this award will ultimately allow researchers to perform genetic modification of cells nearly 10,000 times faster than the current state of the art, while maintaining separation of samples to avoid cross-contamination. This innovation enables continuous flow genetic manipulation of cells in a platform that can be easily automated through integration with liquid handling robots for fast, reliable, and scalable cell engineering.
There are several major highlights and achievements from this effort. With respect to manufacturing, we have used 3D printing to prototype dozens of potential pipette tip designs. As a result we have been able to successfully incorporate a microfluidic channels into pipette tips and achieve genetic transformation efficiencies comparable or higher than the state of the art cuvette based technology. Related to manufacturing, we also have been able to utilize COMSOL Multiphysics to model the electric field and effective flow patterns resulting from our 3D printed designs. This has allowed us to predict the electrical pulse seen by the cells while they flow through the pipette tips. Lastly, we have been able to show that our electroporation system can be used to genetically transform both prokaryotic and eukaryotic cells. In the case of eukaryotic cells we have tested the platform on marine protists, single celled eukaryotic organisms, and found that we can deliver genetic material to organsims that are classically difficult to manipulate. This suggests that the system could have very broad applicability and increases the potential total addressable market for future commercialization efforts.
Last Modified: 06/21/2018
Modified by: Cullen R Buie
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