Polyelectrolyte Multilayer-Mediated Gene Delivery for Semaphorin Signaling Pathway Control
Polyelectrolyte Multilayer-Mediated Gene Delivery for Semaphorin Signaling Pathway Control
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DOI：
10.1002/smll.201000228
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发表时间：
2010-11-05
期刊：
影响因子：
13.3
通讯作者：
Ogier, Joelle
中科院分区：
文献类型：
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作者：
Richard, Doriane;Nguyen, Isabelle;Ogier, Joelle
多层膜（MPFs）的能力，以控制参与一个共同的细胞信号传导活性的两个基因编码细胞受体的顺序表达，同时实现了一个完整的功能信号转导。作为代表以自上而下方式进行的细胞信号传导过程的功能模型系统，选择由信号蛋白3A（Sema 3A）诱导的细胞塌陷作为靶标。聚电解质多层依次用两种质粒功能化，分别编码神经纤毛蛋白-1（NRP-1）和丛蛋白-A1（Plx-A1），作为Sema 3A的共受体。通过使用透明质酸和壳聚糖作为结构组分，用于将质粒DNA层掺入到由聚烯丙胺盐酸盐和聚-4-苯乙烯磺酸钠制成的前体膜上，建立了双链体系统;该系统能够以允许控制两种质粒DNA构建体表达的时间和各自顺序的方式将两种质粒递送至Cos-1细胞。重要的是，观察到在Sema 3A刺激后，共表达Plx-A1和NRP-1的COS-1细胞显示出塌陷表型，这是由多层累积方案确定的，并且嵌入MPF中的两种转基因的表达产物在几天内暂时起作用，同时充当它们的Sema 3A共受体的作用。
The capability of multilayered polyelectrolyte films (MPFs) to control the sequential expression of two genes encoding cell receptors involved in a common cell signalling activity is shown, while achieving a fully functional signal transduction. As a functional model system representative of a cell signalling process that proceeds in a top-down manner, cell collapse induced by semaphorin 3A (Sema3A) was chosen as the target. Polyelectrolyte multilayers were sequentially functionalized with two plasmids encoding Neuropilin-1 (NRP-1) and Plexin-A1 (Plx-A1), respectively, acting as co-receptors for Sema3A. By using hyaluronan and chitosan as structural components for the incorporation of plasmid DNA layers onto precursor films made of poly-allylamine hydrochloride and poly-sodium-4-styrenesulfonate, the polyelectrolyte system is established; this systems is capable of delivering both plasmids to Cos-1 cells in a manner that permits control over the timing and the respective order in which the two plasmid DNA constructs are expressed. Importantly, it was observed that, following Sema3A stimulation, COS-1 cells co-expressing Plx-A1 and NRP-1 display a collapse phenotype, which is determined by the multilayer build-up scheme, and that the expression products of both transgenes embedded in MPFs are temporally functional over several days while acting their role of co-receptors for Sema3A.