BioQuant, Ruprecht-Karls-University Heidelberg, 69120 Heidelberg, Germany.
Life Sciences Center, Vilnius University, Vilnius 10223, Lithuania.
Genome Res. 2017 Oct;27(10):1752-1758. doi: 10.1101/gr.215103.116. Epub 2017 Sep 5.
Delivery of large and functionally active biomolecules across cell membranes presents a challenge in cell biological experimentation. For this purpose, we developed a novel solid-phase reverse transfection method that is suitable for the intracellular delivery of proteins into mammalian cells with preservation of their function. We show results for diverse application areas of the method, ranging from antibody-mediated inhibition of protein function to CRISPR/Cas9-based gene editing in living cells. Our method enables prefabrication of "ready to transfect" substrates carrying diverse proteins. This allows their easy distribution and standardization of biological assays across different laboratories.
将大型生物分子和具有功能的生物分子递送到细胞膜内是细胞生物学实验中的一个挑战。为此,我们开发了一种新颖的固相反向转染方法,该方法适用于将蛋白质递送到哺乳动物细胞内,同时保持其功能。我们展示了该方法在多种应用领域的结果,包括抗体介导的蛋白质功能抑制到活细胞中的 CRISPR/Cas9 基因编辑。我们的方法能够预制携带各种蛋白质的“准备转染”底物。这使得它们能够在不同实验室之间轻松分配和标准化生物测定。