Centro de Investigação em Genética Molecular Humana, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
Nanotechnology. 2010 Dec 17;21(50):505101. doi: 10.1088/0957-4484/21/50/505101. Epub 2010 Nov 22.
The use of gold nanoparticles (AuNPs) has been gaining momentum as vectors for gene silencing strategies, combining the AuNPs' ease of functionalization with DNA and/or siRNA, high loading capacity and fast uptake by target cells. Here, we used AuNP functionalized with thiolated oligonucleotides to specifically inhibit transcription in vitro, demonstrating the synergetic effect between AuNPs and a specific antisense sequence that blocks the T7 promoter region. Also, AuNPs efficiently protect the antisense oligonucleotide against nuclease degradation, which can thus retain its inhibitory potential. In addition, we demonstrate that AuNPs functionalized with a thiolated oligonucleotide complementary to the ribosome binding site and the start codon, effectively shut down in vitro translation. Together, these two approaches can provide for a simple yet robust experimental set up to test for efficient gene silencing of AuNP-DNA conjugates. What is more, these results show that appropriate functionalization of AuNPs can be used as a dual targeting approach to an enhanced control of gene expression-inhibition of both transcription and translation.
金纳米粒子(AuNPs)作为基因沉默策略的载体已经越来越受到关注,因为它将 AuNPs 的易于功能化与 DNA 和/或 siRNA 结合,具有高载量和快速被靶细胞摄取的特点。在这里,我们使用巯基化寡核苷酸功能化的 AuNP 来特异性地抑制体外转录,证明了 AuNP 与特异性反义序列之间的协同作用,该反义序列可以阻断 T7 启动子区域。此外,AuNP 能够有效地保护反义寡核苷酸免受核酸酶的降解,从而保持其抑制潜力。此外,我们还证明了与核糖体结合位点和起始密码子互补的巯基化寡核苷酸功能化的 AuNP 可以有效地抑制体外翻译。总之,这两种方法可以提供一种简单而强大的实验设置,用于测试 AuNP-DNA 缀合物的高效基因沉默。更重要的是,这些结果表明,适当的 AuNP 功能化可以用作双重靶向方法,以增强对基因表达的控制——抑制转录和翻译。