SBIR Phase I: A scalable high-throughput cell engineering platform

这个小型企业创新研究(SBIR)项目的更广泛影响/商业潜力是开发一个可扩展的自动化基因转化平台，速度比目前最先进的平台快10,000倍。合成生物学和基因工程领域目前受到用外来DNA对微生物重新编程的能力的限制。在DNA的合成、基因工程微生物的筛选和生物信息学方面都取得了重大进展。然而，用于运送DNA和进行基因转化的技术并没有以类似的方式取得进展。这项SBIR的第一阶段将产生一个原型高通量遗传转化平台，以展示该系统的实用性。这一系统将使基因工程师能够更快地开发微生物，用于生产生物工程化学品和材料。这个SBIR第一阶段项目建议使用快速、可靠和可扩展的专有流通式电穿孔技术开发一个高通量、自动化的细菌遗传转化平台。细胞基因工程的关键一步是引入外来DNA，对细胞进行重新编程。电穿孔，即使用脉冲电场使细胞通透，是将DNA输送到微生物中的最有效和最广泛的方法。最先进的电穿孔技术包括将细胞和DNA暴露在均匀电场中的试管。然而，目前这一过程缓慢、劳动密集型和昂贵。拟议的技术可以通过增加现有的液体处理机器人来实现自动化，当并行操作时，与目前的方法相比，可以将遗传转化率提高高达10,000倍。这将代表依赖遗传转化领域的范式转变，在这些领域，使用电穿孔传递DNA目前是一个主要瓶颈。最终，目标是满足对高通量基因转化平台的需求，以加速合成生物学的创新。

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.