Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA.
Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1870-5. doi: 10.1073/pnas.0909350107. Epub 2010 Jan 11.
A generalized platform for introducing a diverse range of biomolecules into living cells in high-throughput could transform how complex cellular processes are probed and analyzed. Here, we demonstrate spatially localized, efficient, and universal delivery of biomolecules into immortalized and primary mammalian cells using surface-modified vertical silicon nanowires. The method relies on the ability of the silicon nanowires to penetrate a cell's membrane and subsequently release surface-bound molecules directly into the cell's cytosol, thus allowing highly efficient delivery of biomolecules without chemical modification or viral packaging. This modality enables one to assess the phenotypic consequences of introducing a broad range of biological effectors (DNAs, RNAs, peptides, proteins, and small molecules) into almost any cell type. We show that this platform can be used to guide neuronal progenitor growth with small molecules, knock down transcript levels by delivering siRNAs, inhibit apoptosis using peptides, and introduce targeted proteins to specific organelles. We further demonstrate codelivery of siRNAs and proteins on a single substrate in a microarray format, highlighting this technology's potential as a robust, monolithic platform for high-throughput, miniaturized bioassays.
一个能够将多种生物分子高通量地导入活细胞的通用平台,可以改变人们探测和分析复杂细胞过程的方式。在这里,我们展示了使用表面修饰的垂直硅纳米线对永生和原代哺乳动物细胞进行空间定位、高效和通用的生物分子传递。该方法依赖于硅纳米线穿透细胞膜的能力,随后将表面结合的分子直接释放到细胞质中,从而实现了生物分子的高效传递,而无需化学修饰或病毒包装。这种方式可以评估将广泛的生物效应物(DNA、RNA、肽、蛋白质和小分子)导入几乎任何细胞类型的表型后果。我们表明,该平台可用于使用小分子引导神经元祖细胞生长,通过递送 siRNA 降低转录水平,使用肽抑制细胞凋亡,并将靶向蛋白导入特定细胞器。我们进一步证明了在单个基片上以微阵列格式共递送 siRNA 和蛋白质,突出了该技术作为一种强大的、整体式高通量、微型化生物测定平台的潜力。