Electroporation Knows No Boundaries: The Use of Electrostimulation for siRNA Delivery in Cells and Tissues.
Electroporation Knows No Boundaries: The Use of Electrostimulation for siRNA Delivery in Cells and Tissues.
复制标题
DOI：
10.1177/1087057115579638
复制
发表时间：
2015-09
影响因子：
--
通讯作者：
Ketteler R
中科院分区：
文献类型：
--
作者：
Luft C;Ketteler R
RNA干扰(RNAi)的发现使功能基因组学领域的几项突破性发现成为可能。RNAi技术已成为识别药物靶标的主要工具之一，并已得到稳步改进，可以在细胞系、组织和整个生物体中进行基因操作。在高通量筛选中使用RNAi的主要障碍之一是向细胞和组织输送。一些类型的细胞对标准方法如脂质体或磷酸钙沉淀法的高效转染法无效，需要不同的方法。电穿孔是一种功能强大、用途广泛的方法，可将RNA、DNA、多肽和小分子送入细胞系和原代细胞，以及整个组织和生物体。尤其令人感兴趣的是，在高通量筛选和治疗应用中，使用电穿孔传递小干扰RNA寡核苷酸和成簇的规则间隔的短回文重复/Cas9质粒载体。在这里，我们将回顾电穿孔技术在细胞系和组织高通量筛选中的应用。
The discovery of RNA interference (RNAi) has enabled several breakthrough discoveries in the area of functional genomics. The RNAi technology has emerged as one of the major tools for drug target identification and has been steadily improved to allow gene manipulation in cell lines, tissues, and whole organisms. One of the major hurdles for the use of RNAi in high-throughput screening has been delivery to cells and tissues. Some cell types are refractory to high-efficiency transfection with standard methods such as lipofection or calcium phosphate precipitation and require different means. Electroporation is a powerful and versatile method for delivery of RNA, DNA, peptides, and small molecules into cell lines and primary cells, as well as whole tissues and organisms. Of particular interest is the use of electroporation for delivery of small interfering RNA oligonucleotides and clustered regularly interspaced short palindromic repeats/Cas9 plasmid vectors in high-throughput screening and for therapeutic applications. Here, we will review the use of electroporation in high-throughput screening in cell lines and tissues.