Kalies Stefan, Heinemann Dag, Schomaker Markus, Murua Escobar Hugo, Heisterkamp Alexander, Ripken Tammo, Meyer Heiko
Laser Zentrum Hannover e.V., Hollerithallee 8, 30419 Hannover, Germany.
J Biophotonics. 2014 Oct;7(10):825-33. doi: 10.1002/jbio.201300056. Epub 2013 Jun 6.
Several cell transfection techniques have been developed in the last decades for specific applications and for various types of molecules. In this context, laser based approaches are of great interest due to their minimal invasiveness and spatial selectivity. In particular, laser induced plasmon based delivery of exogenous molecules into cells can have great impact on future applications. This approach allows high-throughput laser transfection by excitation of plasmon resonances at gold nanoparticles non-specifically attached to the cell membrane. In this study, we demonstrate specific gene-knockdown by transfection of Morpholino oligos using this technique with optimized particle size. Furthermore, we evaluated the cytotoxicity of plasmonic laser treatment by various assays, including LDH activity and ROS formation. In summary, this study gives important insights into this new approach and clearly demonstrates its relevance for possible biological applications.
在过去几十年中，已经开发出了几种针对特定应用和各类分子的细胞转染技术。在这种背景下，基于激光的方法因其微创性和空间选择性而备受关注。特别是，基于激光诱导等离子体将外源性分子递送至细胞的方法可能会对未来的应用产生重大影响。这种方法通过激发非特异性附着在细胞膜上的金纳米颗粒的等离子体共振来实现高通量激光转染。在本研究中，我们使用优化粒径的该技术通过转染吗啉代寡核苷酸来证明特异性基因敲低。此外，我们通过包括乳酸脱氢酶（LDH）活性和活性氧（ROS）形成在内的各种测定方法评估了等离子体激光处理的细胞毒性。总之，本研究为这种新方法提供了重要见解，并清楚地证明了其在可能的生物学应用中的相关性。
