Department of Medicine, Health Sciences, University of California, San Diego, La Jolla, CA 92093, USA; Department of Molecular Biology, Biological Sciences, University of California, San Diego, La Jolla, CA 92093, USA.
Department of Medicine, Health Sciences, University of California, San Diego, La Jolla, CA 92093, USA; Department of Molecular Biology, Biological Sciences, University of California, San Diego, La Jolla, CA 92093, USA; Biological Sciences Graduate Program, University of California, San Diego, La Jolla, CA 92093, USA.
STAR Protoc. 2024 Sep 20;5(3):103107. doi: 10.1016/j.xpro.2024.103107. Epub 2024 Jul 3.
Electroporation is a technique to introduce DNA constructs into cells using electric current. Here, we present a protocol to electroporate DNA plasmids into Ciona robusta embryos at the 1-cell stage. We describe steps for setting up and conducting electroporation. We then detail procedures for collecting, fixing, and mounting embryos and counting expression. This protocol can be used to study the expression of enhancers via reporter assays, manipulating cells using genes or modified genes such as dominant negatives, and genome editing. For complete details on the use and execution of this protocol, please refer to Song, et al..
电穿孔是一种使用电流将 DNA 构建体导入细胞的技术。在这里，我们介绍了一种在 1 细胞期将 DNA 质粒电穿孔到海鞘胚胎中的方案。我们描述了设置和进行电穿孔的步骤。然后，我们详细介绍了收集、固定和安装胚胎以及计数表达的程序。该方案可用于通过报告基因检测研究增强子的表达，使用基因或修饰基因（如显性负突变体）操纵细胞，以及基因组编辑。有关该方案使用和执行的完整详细信息，请参阅 Song 等人的文章。
