SBIR Phase I: Non-templated Enzymatic Synthesis of Polydeoxynucleotides

该小型企业创新研究（SBIR）第一阶段提案要求提供资金，以开发一种用于分子，系统和合成生物学应用的单链DNA分子的受控非模板酶促合成的新技术。当实施时，该技术代表了自1981年引入亚磷酰胺介导的固体支持物方法以来寡脱氧核苷酸和多脱氧核苷酸生产方式的最显著变化。新工艺将通过降低成本，将直接合成的DNA长度增加10倍至50倍，将周转时间从数周和数月减少到数天，并消除大量有毒废物的产生，从而缓解分子和合成生物学工作流程中的瓶颈。将使用一种独特的酶，其催化脱氧核苷酸的快速聚合而不需要模板分子。该项目将创造新的类似物，这些类似物是合适的酶底物，并允许逐步添加核苷酸，模拟金标准化学合成方法，但在一个更简单的水相酶促过程中。该项目的更广泛的影响/商业潜力，如果成功，将是创造下一代DNA合成方法。 这种能力不仅将加强当前的研究，而且将使生物医学，计算机科学，纳米光电子学和生物纳米技术等多学科研究领域不断发展。利用DNA纳米结构进行药物递送和单细胞体内分析的研究正在发展。DNA被用作信息存储介质和计算本身。工程师们正在研究基因电路作为电子元件的生物模拟物，如振荡器和晶体管。这个最有趣、最复杂、最重要的生物分子肯定有许多我们无法想象的应用。正如25年前第一代DNA合成技术开启了生物技术的新时代一样，这里介绍的能力将开启新一代生物应用，并有可能满足社会未来的需求。

This Small Business Innovation Research (SBIR) Phase I proposal requests funding to develop a novel technology for the controlled non-templated enzymatic synthesis of single strand DNA molecules for use in molecular, systems, and synthetic biology applications. When implemented, this technology represents the most significant change to the way that oligodeoxynucleotides and polydeoxynucleotides have been produced since the introduction of the phosphoramidite mediated solid support method in 1981. The new process will relieve the bottle necks in molecular and synthetic biology workflows by reducing costs, increasing the lengths of DNA directly synthesized by 10x to 50x, reducing turnaround times from weeks and months to days, and eliminating production of tons of toxic wastes. A unique enzyme will be used that catalyzes the rapid polymerization of deoxynucleotides without need of a template molecule. The project will create novel analogs that are suitable enzymatic substrates and allow the stepwise addition of nucleotides mimicking the gold standard chemical synthesis method but in a simpler and aqueous enzymatic process.The broader impact/commercial potential of this project, if successful, will be to create a next generation approach to DNA synthesis. Such capabilities will not only enhance current research, but will enable evolving multidisciplinary research areas such as biomedicine, computer science, nano-optoelectronics, and bionanotechnology. Research utilizing DNA nanostructures for drug delivery and single cell in-vivo analysis is evolving. DNA is being used as an information storage medium and for computing itself. Engineers are investigating gene circuits as biological analogs of electronic components such as oscillators and transistors. This most interesting, complex and biologically important molecule surely has many applications that cannot yet be imagined. Just as the first generation DNA synthesis technology enabled a new era of biotechnology 25 years ago, the capabilities presented here will enable a new generation of biological applications and potentially provide for society's future needs.