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

随着新的基于DNA的疗法和技术的发展，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难的序列的需要已经使DNA的生产达到了极限。目前的DNA生产方法依赖于细菌发酵，这在细菌生长过程中存在生产能力问题和低序列稳定性。Touchlight的合成生物学DNA生产工艺克服了目前使用酶法生产细菌发酵的局限性。这种体外DNA技术能够使用DNA聚合酶扩增DNA模板和原核端粒酶切割扩增的DNA并将其连接到最小盒中以用于治疗和合成生物学应用，从而产生大规模、高质量的构建体。该方法的不断发展已经导致DNA制造的能力增加和足迹减少，缓解了与核酸生产相关的一些当前瓶颈。尽管在该方法中已经在DNA制造中取得了很大的进步，但对DNA在细胞环境中如何表现的更好理解将允许生产改进的DNA产物和平台。和核靶向能力，导致蛋白质表达增加，然而，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了解对于持续改进Touchlight的核心结构至关重要。该项目产生的知识将提高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转化改进将有助于满足工业合作伙伴对DNA构建体日益增长的需求，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.