Surface- and hydrogel-mediated delivery of nucleic acid nanoparticles.
Surface- and hydrogel-mediated delivery of nucleic acid nanoparticles.
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DOI：
10.1007/978-1-62703-140-0_11
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Segura, Tatiana
中科院分区：
文献类型：
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作者：
Pannier, Angela K;Segura, Tatiana
细胞群体内的基因表达可以通过基因递送方法直接改变。传统上，对于非病毒递送，编码或靶向感兴趣基因的质粒或siRNA分子被包装在纳米颗粒内。然后将这些纳米颗粒递送到接种在组织培养塑料上的细胞周围的培养基中;这种技术称为推注递送。虽然推注递送被广泛用于筛选有效的递送载体和研究体外基因功能，但这种递送策略可能不会导致所有细胞类型的有效基因转移，或者可能无法鉴定那些在体内有效的递送载体。此外，推注递送不能用于需要基因表达模式化的应用中。在本章中，我们描述了将材料表面（即，表面介导的递送）或水凝胶支架（即，水凝胶介导的递送）以有效地递送基因。本章包括协议表面介导的DNA传递集中在最简单和最有效的方法，其中包括非特异性固定的DNA复合物（聚合物和脂质载体）到血清包被的细胞培养聚苯乙烯和自组装单分子层的烷硫醇金。此外，描述了将DNA/阳离子聚合物纳米颗粒包封到水凝胶支架中的方案，包括用于包封少量DNA（0.2 μ g/穆尔）的方法，因为大量DNA的掺入由于聚集而造成显著的挑战。
Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for efficient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in efficient gene transfer for all cell types or may not identify those delivery vehicles that will be efficient in vivo. Furthermore, bolus delivery cannot be used in applications where patterning of gene expression is needed. In this chapter, we describe methods that incorporate material surfaces (i.e., surface-mediated delivery) or hydrogel scaffolds (i.e., hydrogel-mediated delivery) to efficiently deliver genes. This chapter includes protocols for surface-mediated DNA delivery focusing on the simplest and most effective methods, which include nonspecific immobilization of DNA complexes (both polymer and lipid vectors) onto serum-coated cell culture polystyrene and self-assembled monolayers of alkanethiols on gold. Also, protocols for the encapsulation of DNA/cationic polymer nanoparticles into hydrogel scaffolds are described, including methods for the encapsulation of low amounts of DNA (0.2 mug/muL) since incorporation of high amounts of DNA poses significant challenges due to aggregation.