Synthesis of Modified Nucleotide Triphosphates for Enzymatic DNA Synthesis**

DNA测序技术在10年内将阅读DNA的价格降低了1万多次。合成生物学的瓶颈现在是写入(即合成)DNA。三个主要经济体(美国、法国、英国)的初创公司正在研究下一代酶促DNA合成技术，以取代目前的化学技术。Spindle Biotech仅专注于开发加拿大的酶促DNA合成技术。目前，唯一在商业上可行的方法是20世纪70年代开发的两步法。这种方法化学合成短DNA片段，并在一个容易出错且耗时的步骤中组装它们。这一过程花费的时间长一个数量级，估计每年产生1200,000升危险化学废物，包括乙腈、三氯乙酸、甲苯、四氢呋喃和吡啶。酶DNA合成是降低长合成DNA成本和周转时间的唯一方法，它有助于加速疫苗生产，设计能够限制癌症生长的新型酶，生产可持续的生物燃料，提高作物产量，并检测和分解土壤、空气和水中的环境污染物。**拟议的创新是一组16个受保护的脱氧核糖核酸三磷酸盐，对应于四个天然核苷酸的每个排列和从文献中选择的四个保护基团。这些核苷酸将在多伦多大学斯卡伯勒分校合成。这些核苷酸将由Spindle Biotech Inc.在多伦多市中心的JLABS孵化器中进行筛选。针对酶的核苷酸文库和反应条件，优化DNA合成过程。已经确定了一些拟议合成的起点。工业合作伙伴Spindle Biotech Inc.是一家加拿大独资公司，专门致力于开发加拿大的酶DNA合成技术。这个项目中设计的核苷酸将被并入正在并行开发的DNA合成器中。其影响是深远的，包括公共卫生和国家安全。

DNA sequencing technology has dropped the price of reading DNA over 10,000 times in 10 years. The bottleneck of synthetic biology is now writing (i.e. synthesizing) DNA. Startups in three major economies (US, France, UK) are working on next-generation enzymatic DNA synthesis technology to replace the current chemical technology. Spindle Biotech is solely focused on developing Canadian technology for enzymatic DNA synthesis. Currently, the only commercially viable method is a two-step process developed in the 1970s. This method chemically synthesizes short DNA fragments and assembles them in an error-prone and time-consuming step. This process takes an order of magnitude longer and is responsible for an estimated 1,200,000 litres of hazardous chemical waste per year, including acetonitrile, trichloroacetic acid, toluene, tetrahydrofuran, and pyridine. Enzymatic DNA synthesis is the only way to reduce the cost and turnaround time for long synthetic DNA, which helps in accelerating vaccine production, engineering novel enzymes that have the ability to restrict cancer growth, producing sustainable biofuels, improving crop yield, and detecting and breaking down environmental pollutants in the soil, air, and water.**The proposed innovation is a set of sixteen protected deoxyribonucleotide triphosphates, corresponding to the each permutation of the four natural nucleotides with four protecting groups chosen from literature. The nucleotides will be synthesized at the University of Toronto, Scarborough. The nucleotides will be screened by Spindle Biotech Inc. at the JLABS incubator in downtown Toronto. Nucleotides against a library of enzymes and reaction conditions to optimize the DNA synthesis process. A number of starting points for the proposed syntheses have been identified. The industrial partner, Spindle Biotech Inc., is a Canadian-owned company solely focused on developing Canadian technology for enzymatic DNA synthesis. The nucleotides designed in this project will be incorporated into a DNA synthesizer being developed in parallel. The implications are far reaching, including public health and national security.