Electroporation in biology: methods, applications, and instrumentation.
Electroporation in biology: methods, applications, and instrumentation.
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DOI：
10.1016/0003-2697(88)90035-8
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发表时间：
1988-11
影响因子：
2.9
通讯作者：
Huntington Potter
中科院分区：
文献类型：
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作者：
Huntington Potter
电穿孔技术-通过高压电击在细胞膜上形成孔或小孔-在生物学中得到了广泛的应用。首先用于诱导细胞通过它们的质膜融合（（1 -4）;综述见（5）），然后Neumann和他的同事发现电穿孔（6，7）允许小鼠成纤维细胞（L细胞）摄取和表达外源DNA。然而，由于L细胞很容易通过传统的基因转移方法来摄取DNA，例如，通过摄取磷酸钙/DNA共沉淀物（8），因此最初并不清楚新方法是否可以应用于其他细胞类型。然后在骨髓瘤细胞（6，9）和神经元细胞系（10）中证明了电穿孔。我们扩展并修改了电穿孔（9）以允许将外源DNA引入到广谱的细胞类型中，包括例如淋巴细胞、神经元细胞、内分泌细胞、原代动物细胞、肝细胞瘤细胞、造血干细胞、单细胞生物体、植物原生质体和细菌（参见例如（lo-19））。事实上，电穿孔产生高频率的永久转染子，具有高效率的瞬时基因表达，并且比替代技术更容易进行，这导致其在许多应用中的使用越来越多。事实上，在过去的几年里，电穿孔已经从一些发育实验室中脱颖而出，成为许多情况下基因转移的首选方法（见图1）。
The technique of electroporation-the formation of holes or pores in the cell membrane by high voltage electric shock-has found widespread application in biology. First used to induce cells to fuse via their plasma membranes ((l-4); for review, see (5)), electroporation was then found by Neumann and his colleagues (6, 7) to allow mouse fibroblasts (L cells) take up and express exogenous DNA. However, because L cells are easily made to take up DNA by the traditional methods of gene transfer, for instance, by uptake of calcium phosphate/DNA coprecipitates (8), it was not at first clear whether the new procedure could be applied to other cell types. Electroporation was then demonstrated in myeloma cells (6, 9) and a neuronal cell line (10). We extended and modified electroporation (9) to allow the introduction of exogenous DNA into a broad spectrum of cells types, including, for example, lymphocytes, neuronal cells, endocrine cells, primary animal cells, hepatoma cells, hematopoietic stem cells, unicellular organisms, plant protoplasts, and bacteria (see, for example,(lo-19)). The fact that electroporation yields a high frequency of permanent transfectants, has a high efficiency of transient gene expression, and is substantially easier to carry out than alternative techniques has resulted in its increasing use in many applications. Indeed, in the last few years, electroporation has moved out of a few developmental laboratories to become the method of choice for gene transfer in many situations (see Fig. 1).