Efficient and Selective Delivery of Oligonucleotides

通过基因修饰调节细胞行为的能力作为一种治疗策略具有巨大的潜力，并为阐明基因功能提供了强大的工具（所谓的功能基因组学）。 反义寡核苷酸，最常见的是长度为15-25个核苷酸的单链DNA分子，通过与来自靶基因的mRNA上的互补片段结合来调节基因表达。 虽然反义技术正在成为功能基因组学的可行治疗实体和平台，但其广泛应用的主要障碍仍然存在：以生物学有效的量和功能完整但无毒的形式将遗传物质（多核酸）递送至细胞。我们的总体目标是将反义分子选择性地递送到靶细胞类型（肝细胞），从而导致感兴趣的基因表达的低剂量抑制。 为了实现这一目标，我们将开发一个新的家庭的多功能DNA传递载体（多路复用）使用组合合成方法。 这些载体将具有使DNA缩合的仿生聚合物、使细胞膜不稳定的阳离子肽和使它们靶向肝细胞（原代肝细胞）的半乳糖部分。 将表征载体的大小、稳定性和细胞毒性。 我们将研究这些材料对靶细胞与非靶细胞的吸附，并在胶体化学数学模型的框架中解释结果，该模型将用于改进和优化载体的组成。 这些载体的有效性，提供基因表达调节反义寡核苷酸将进行评估和评估，以进一步完善的方法。 我们期望长期的结果是一个选择性和有效的方法寡核苷酸输送治疗和功能基因组学的应用。

The ability to modulate cell behavior through genetic modification has great potential as a therapeutic strategy, as well as providing a powerful tool for elucidating gene function (so-called functional genomics). Antisense oligonucleotides, which are most commonly single-stranded DNA molecules 15-25 nucleotides in length, modulate gene expression by binding to a complementary segment on the mRNA from the target gene. While antisense technology is becoming a viable therapeutic entity and platform for functional genomics, a major barrier to its widespread practice still exists: the delivery of the genetic material (polynucleic acid) to cells in a quantity that is biologically effective and in a form that is functionally intact, yet non-toxic. Our overall goal is to deliver antisense molecules selectively to a target cell type (hepatocytes), resulting in low-dose inhibition of expression of genes of interest. To achieve this goal, we will develop a new family of multifunctional DNA delivery vectors (multiplexes) using a combinatorial synthesis approach. These vectors will possess biomimetic polymers that condense DNA, cationic peptides that destabilize cellular membranes, and galactose moieties that target them to hepatocytes (primary liver cells). Vectors will be characterized for size, stability, and cytotoxicity. We will study the adsorption of these materials to target vs. non-target cells, and interpret the results in the framework of a colloid-chemical mathematical model, which will be used to refine and optimize the composition of the vectors. The effectiveness of these vectors to deliver gene expression-modulating antisense oligonucleotides will be evaluated and assessed to further refine the approach. W expect the long-term outcome to be a selective and efficient method for oligonucleotide delivery for therapeutic and functional genomics applications.