Novel Virus-Like Particles for Nuclear DNA Delivery

一种介于经典病毒和非病毒基因传递领域之间的新兴混合体：合成病毒样载体已经取得了进展。这些浓缩的DNA颗粒携带一种或多种病毒样特征，使它们在某些传递步骤中更有效，同时保持比真正的病毒的优势。这些研究的第一个目标是为构建这种病毒样颗粒的新家族奠定基础。我们计划使用各种合成聚合物和传统聚合物以及各种化学方法来形成具有正、负和近中性表面电位的凝聚和共价交联（笼状）DNA颗粒。然后，重点将转移到增强DNA核运输，这是成功的基因传递的最不可克服的障碍之一。这些颗粒在DNA核递送中的效率将在微注射细胞中进行测试，以测试是否有共价附着的核靶向信号。根据标记DNA的亚细胞定位和标记基因表达的定量（至少比对照增加10-20倍），将选择最有效的颗粒配制技术。然后可以调整颗粒以适应许多不同输送系统的需要。拟议的商业应用：在拟议的研究中开发的核转运技术将与细胞受体配体和内溶剂结合到基因治疗载体中。鉴于非病毒载体行业意识到低效率的核运输阻碍基因成功传递的力量，一种克服这一障碍的新技术将具有巨大的商业价值。

An emerging hybrid between the classical viral and non-viral gene delivery fields: synthetic virus-like vectors have gained ground. These condensed DNA particles carry one or more virus-like characteristics that make them more efficient at (a) certain step(s) of delivery, yet maintaining advantages over real viruses. The first goal of these studies is to lay the foundation for the construction of a novel family of such virus-like particles. We plan to use a variety of synthetic and traditional polymers and a variety of chemical approaches to form condensed and covalently cross-linked (caged) DNA particles with positive, negative and near-neutral surface potentials. Then the emphasis will be shifted to enhancing DNA nuclear transport, which is one of the most invincible obstacles to successful gene delivery. The efficiency of these particles in DNA nuclear delivery will be tested without or with covalently attached nuclear targeting signals, in microinjected cells. Based on the sub-cellular localization of labeled DNA and on the quantitation of marker gene expression (minimum 10-20x increase over control) the most efficient particle formulation technology(ies) will be selected. The particles then can be adjusted to the needs of many different delivery systems. PROPOSED COMMERCIAL APPLICATION: The nuclear transport technology developed in the proposed studies will be incorporated into gene therapy vectors with ligands for cellular receptors and with endosomolytic agents. Given that the non-viral vector industry realizes the power of inefficient nuclear transport to obstruct successful gene delivery, a new technology for overcoming this hurdle will have great commercial value.