Lipid-Based Nanoparticles that Controllably Synthesize Proteins

将蛋白质和RNA运送到特定的细胞和组织是一个重要的问题，因为这个问题的工程解决方案将应用于治疗疾病和理解基本的生物过程。已经开发的用于输送小分子的技术，如靶向癌症的纳米颗粒和药物洗脱支架，已经在商业上取得了成功。然而，由于生物大分子在体内的不稳定性、易降解性和引发免疫反应的潜力，递送生物大分子已被证明更具挑战性，因此，尚未达到全部商业潜力。目前正在开发的材料和技术旨在稳定和屏蔽活性生物分子。我的项目将探索一种完全不同的方法——只在所需的时间和地点合成活性生物分子。我们将在这个项目中回答的主要问题是：蛋白质和RNA能否在脂质纳米囊泡内可控地表达？我们的主要目标是开发纳米颗粒，这些纳米颗粒最初是惰性的，但当被光激活时，将合成具有功能的生物分子，用于一系列应用。我是一个团队的一员，该团队最近展示了大肠杆菌细胞提取物（一种常用的蛋白质表达系统）和DNA模板可以被封装在脂质囊泡内，从而产生可以合成模型蛋白质的纳米颗粒。我的项目的具体目标是设计这个系统，以i)增加可以合成的蛋白质的产量和范围，ii)为RNAi合成功能性小RNA，以及iii)开发用于筛选蛋白质库的高通量微流控系统。该计划的影响将是一种新的生化技术和药物输送技术的发展。它可以帮助实现蛋白质和RNA输送的全部商业潜力，从而使加拿大受益。该计划还将通过培养高素质的创业人才使加拿大受益。学员将在协作环境中接触基础科学和工程，同时学习一系列应用于及时解决问题的最先进技术。

Delivering proteins and RNA to specific cells and tissues is an important problem because engineering solutions to this problem will have applications for treating diseases and understanding fundamental biological processes. The technologies that have been developed to deliver small molecules, such as cancer-targeting nanoparticles and drug-eluting stents, have been commercially successful. However delivering large biomolecules has proven more challenging, due to their instability in vivo, susceptibility to degradation, and potential to elicit immune responses, and consequently, the full commercial potential has not been reached. The materials and technologies being developed to-date aim to stabilize and shield active biomolecules. My program will explore a fundamentally different approach - to synthesize the active biomolecules only in desired locations at desired times. The main question we will answer in this program is: Can proteins and RNA be controllably expressed inside of lipid nano-vesicles? Our main objective is to develop nanoparticles that are initially inert, but when activated by light will synthesize functional biomolecules for a range of applications. I was part of a team that recently showed that E. coli cell extract (a commonly used protein-expression system) and DNA templates can be encapsulated inside of lipid vesicles, creating nanoparticles that could synthesize model proteins. The specific objectives of my program are to engineer this system to i) increase both the yield and the range of proteins that can be synthesized, ii) synthesize functional small RNA for RNAi, and iii) develop a high-throughput microfluidic system for screening protein libraries. The impact of this program will be the development of a new biochemical technique and drug-delivery technique. It can benefit Canada by helping achieve the full commercial potential of protein and RNA delivery. The program will also benefit Canada by training highly qualified entrepreneurial personnel. Trainees will be exposed to fundamental science and engineering in a collaborative environment while learning a range of state-of-the-art techniques applied to timely problems.