Institute of Toxicology, Johannes-Gutenberg University of Mainz, Obere Zahlbacher St. 67, 55131 Mainz, Germany.
STAR Protoc. 2022 Sep 16;3(3):101512. doi: 10.1016/j.xpro.2022.101512. Epub 2022 Jul 1.
Genetic silencing of leukemia-associated proteins with small interfering RNAs (siRNAs) is a straightforward way to delineate their functions. It can be very challenging to deliver siRNAs to leukemia-derived cells with high transfection efficiency and without compromising their viability. This protocol describes an efficient approach to silence oncogenic feline McDonough sarcoma (FMS)-like tyrosine kinase-3 in leukemia cells using siRNAs that are delivered by electroporation. The protocol maintains high cell viability and is generally useful to decrease RNAs encoding proteins of interest. For complete details on the use and execution of this protocol, please refer to Beyer et al. (2022).
利用小干扰 RNA(siRNA)对白血病相关蛋白进行基因沉默是阐明其功能的一种直接方法。用高转染效率且不影响细胞活力的方法将 siRNA 递送到白血病衍生细胞中极具挑战性。本方案描述了一种使用电穿孔将 siRNA 递送至白血病细胞中以沉默致癌性猫 McDonough 肉瘤(FMS)样酪氨酸激酶-3 的有效方法。该方案保持高细胞活力,通常可用于降低编码感兴趣蛋白的 RNA。有关该方案使用和执行的完整详细信息,请参阅 Beyer 等人(2022 年)。