| NSF Org: |
TI Translational Impacts |
| Recipient: |
|
| Initial Amendment Date: | December 21, 2017 |
| Latest Amendment Date: | December 21, 2017 |
| Award Number: | 1747096 |
| Award Instrument: | Standard Grant |
| Program Manager: |
Ruth Shuman
rshuman@nsf.gov (703)292-2160 TI Translational Impacts TIP Directorate for Technology, Innovation, and Partnerships |
| Start Date: | January 1, 2018 |
| End Date: | December 31, 2018 (Estimated) |
| Total Intended Award Amount: | $225,000.00 |
| Total Awarded Amount to Date: | $225,000.00 |
| Funds Obligated to Date: |
|
| History of Investigator: |
|
| Recipient Sponsored Research Office: |
750 MAIN ST CAMBRIDGE MA US 02139-3544 (443)799-3072 |
| Sponsor Congressional District: |
|
| Primary Place of Performance: |
10 Rogers Street Cambridge MA US 02142-1251 |
| Primary Place of
Performance Congressional District: |
|
| Unique Entity Identifier (UEI): |
|
| Parent UEI: |
|
| NSF Program(s): | SBIR Phase I |
| Primary Program Source: |
|
| Program Reference Code(s): |
|
| Program Element Code(s): |
|
| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.084 |
ABSTRACT
The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is the development a scalable, automated, genetic transformation platform that is 10,000X faster than the current state-of-the-art. The fields of synthetic biology and genetic engineering are currently limited by the ability to re-program microorganisms with foreign DNA. There have been significant advances in the synthesis of DNA, screening of genetically engineered microorganisms, and bioinformatics. However, the technology used to deliver DNA and perform genetic transformation has not advanced in a similar way. Phase I of this SBIR will result in a prototype high-throughput genetic transformation platform to demonstrate the utility of the system. This system will allow genetic engineers to more rapidly develop microorganisms for the production of bioengineered chemicals and materials.
This SBIR Phase I project proposes to develop a high-throughput, automated platform for genetic transformation of bacteria using a proprietary flow-through electroporation technology that is fast, reliable, and scalable. A key step in genetic engineering of cells is to introduce the foreign DNA that re-programs the cell. Electroporation, cell permeabilization using pulsed electric fields, is the most efficient and widespread method to deliver DNA into microorganisms for this application. State-of-the-art electroporation involves cuvettes that expose the cells and DNA to uniform electric fields. However, this process is currently slow, labor-intensive, and expensive. The proposed technology can be automated by augmenting existing liquid handling robots, and, when operated in parallel, may improve the genetic transformation rate by up to 10,000X compared to current methods. This will represent a paradigm shift in areas dependent upon genetic transformation where DNA delivery using electroporation is currently a major bottleneck. Ultimately, the goal is to address the need for a high-throughput genetic transformation platform to accelerate innovation in synthetic biology.
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.
Kytopen is a technology company developing an efficient method to deliver genetic materials to cells. Kytopen has developed a proprietary transfection technology known as FlowfectTM which can integrate into automated liquid handling systems. FlowfectTM couples microfluidics with electric fields to reversibly open pores and deliver external material to a diverse array of cell types. In this NSF Phase I SBIR project we sought to develop proof of concept FlowfectTM systems. There were four major objectives for this project. The first objective was the design and testing of prototype FlowfectTM pipette tips. The technology is coupled with automated liquid handling systems by being integrated into pipette tips. Over the course of this project we designed a version of FlowfectTM tips that are compatible with high volume, low cost manufacturing techniques. The second objective for the project was to integrate electrical connections into the microfluidic pipette tips. This was achieved by re-designing the pipette tips to expose the electrodes such that they could be easily connected to a power supply for electrical pulse delivery. The third objective for the project was to automate a single pipette with fluid handling. We exceeded this objective because by the end of the Phase I project we were able to demonstrate automation of eight FlowfectTM pipettes simultaneously. The fourth and final objective was to secure a pilot study with a strategic partner to quantify workflow improvement. By the end of the project we had one pilot study signed and a second pilot study under negotiation with two public biotech companies seeking to commercialize cell therapies.
Over the course of this project our world class team has been able to manufacture and successfully electroporate multiple types of mammalian cells including Jurkat E6-1 T cells and primary human T cells, all with the Flowfect? platform in house. We have completed the design and optimization of the FlowfectTM tips for transfection in T cells by experimentally studying known parameters that have the most significant impact on electroporation. At this phase of the FlowfectTM development all of the devices have been 3D printed to facilitate rapid prototyping. Moreover, through collaborations with industry partners we have gained access to a JANUS G3 BioTx Pro liquid handling system for technological development of the Flowfect? devices.
The rapid, high throughput, and automated engineering of human cells will impact several scientific disciplines of commercial importance. Applications of this technology range from fundamental research in cell physiology to the discovery of new targets for cellular therapies. The applications in cell therapies alone can contribute to a growing multi-billion-dollar industry. The current state of the art in genetic manipulation at the research scale is manually intensive and difficult to incorporate with automated liquid handling systems. The work completed in this project advances a novel system for cell engineering that easily integrates with a diverse array of liquid handling platforms. This will allow researchers in academia and industry to quickly explore a wide array of questions related to gene and cell therapies. Ultimately this technological advance will facilitate research scale cell engineering thousands of times faster than the state of the art, leading to life changing discoveries in the biological sciences and healthcare.
Last Modified: 02/11/2019
Modified by: Paulo A Garcia
Please report errors in award information by writing to: awardsearch@nsf.gov.
