Measurement validation of novel nuclear-targeting DNA as next generation therapies

随着基于DNA的新疗法和技术的发展，对核酸产品的需求持续增加。对大量高质量DNA的需求，以及为先进疗法生产更困难的序列的需要，使DNA的生产达到了极限。目前的DNA生产方法依赖于细菌发酵，这既存在制造能力问题，也存在细菌生长过程中序列稳定性较低的问题。Touchlight的合成生物DNA生产工艺克服了目前使用酶制剂进行细菌发酵的局限性。这种体外DNA技术能够生产大规模、高质量的构建物，使用DNA聚合酶来放大DNA模板，使用原端粒酶来裂解和连接扩增的DNA到最小的盒中，用于治疗和合成生物学应用。这一过程的不断发展导致DNA制造能力的提高和占地面积的减少，缓解了目前与核酸生产相关的一些瓶颈。尽管在DNA制造过程中取得了很大进展，但对DNA在细胞环境中行为的更深入了解将使改进的DNA产品和平台的生产成为可能。Touchlight开发了具有更强的细胞和核靶向能力的新DNA结构，从而增加了蛋白质表达，然而，有限的知识和在细胞内监控DNA的能力使进一步的优化面临挑战。因此，对DNA亚细胞定位的更详细了解对于Touchlight核心结构的持续改进至关重要。该项目产生的知识将增加Touchlight满足DNA需求的能力，并将直接有助于改善人类健康。Touchlight领先的体外DNA制造工艺在核进入效率方面的转化改进将有助于满足工业合作伙伴对DNA结构日益增长的需求，并降低DNA疗法的成本。

With the development of new DNA-based therapies and technologies the demand for nucleic acid products continues to increase. The requirement for large quantities of high-quality DNA and the need to produce more difficult sequences for advanced therapeutics have brought the production of DNA to its limits. Current methods of DNA manufacture rely on bacterial fermentation which suffers from both manufacturing capacity issues and low sequence stability during bacterial growth.Touchlight's synthetic biology DNA production process overcomes the current limitation of bacterial fermentation using enzymatic means of production. This _in vitro_ DNA technology is able to produce large-scale, high-quality constructs using a DNA polymerase to amplify DNA templates and a protelomerase to cleave and ligate amplified DNA into minimal cassettes for use in therapeutics and synthetic biology applications. Continuous developments in the process have resulted in both increased capacity and reduced footprint for DNA manufacture alleviating some of the current bottlenecks associated with nucleic acid productionAlthough large advances in this process have been made in DNA manufacturing a greater understanding of how DNA behaves in a cellular context will allow the production of improved DNA products and platform.Touchlight has developed new DNA constructs with improved cellular and nuclear targeting abilities resulting in increased protein expression however the limited knowledge and ability to monitor DNA within a cell makes further optimisation challenging. A more detailed understanding of the DNA subcellular localisation is therefore essential for the continuing improvement of Touchlight's core constructs.The knowledge generated from this project will increase Touchlight's ability to satisfy DNA demand and will contribute directly to improving human health. Translated improvements in nuclear entry efficiency for Touchlight's leading _in vitro_ DNA manufacturing process will help meet the increasing demand of DNA constructs from Industrial partners and decrease the costs of DNA therapeutics.