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

随着新的基于dna的治疗方法和技术的发展，对核酸产品的需求不断增加。对大量高质量DNA的需求，以及为先进的治疗方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制造方法依赖于细菌发酵，在细菌生长过程中存在制造能力问题和低序列稳定性问题。Touchlight的合成生物学DNA生产工艺克服了目前使用酶生产方法进行细菌发酵的限制。这种体外DNA技术能够使用DNA聚合酶来扩增DNA模板，使用端粒前酶来切割和连接扩增的DNA到最小的磁带中，用于治疗学和合成生物学应用。这一过程的不断发展导致了DNA制造能力的增加和足迹的减少，缓解了目前与核酸生产相关的一些瓶颈。尽管这一过程在DNA制造方面取得了很大进展，但对DNA在细胞环境中的行为的更深入了解将允许生产改进的DNA产品和平台。Touchlight开发了新的DNA结构，具有改进的细胞和核靶向能力，导致蛋白质表达增加，但是有限的知识和监测细胞内DNA的能力使得进一步优化具有挑战性。因此，对DNA亚细胞定位的更详细的了解对于Touchlight核心结构的持续改进至关重要。从这个项目中产生的知识将提高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改进将有助于满足工业合作伙伴对DNA构建物日益增长的需求，并降低DNA治疗的成本。

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.