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胚胎早期发育过程中质粒DNA和吗啉代寡核苷酸在特定组织中的卵内电穿孔技术

In Ovo Electroporation of Plasmid DNA and Morpholinos into Specific Tissues During Early Embryogenesis.

作者信息

McLennan Rebecca, Kulesa Paul M

机构信息

Stowers Institute for Medical Research, Kansas City, MO, USA.

Department of Anatomy and Cell Biology, University of Kansas School of Medicine, Kansas City, KS, USA.

出版信息

Methods Mol Biol. 2019;1976:71-82. doi: 10.1007/978-1-4939-9412-0_6.

Abstract

In ovo electroporation enables transfection of non-viral plasmid DNA and/or morpholinos to fluorescently label and/or perturb gene function in cells of interest. However, targeted electroporation into specific subregions of the embryo can be challenging due to placement and size limitations of the electrodes. Here we describe the basic techniques for in ovo electroporation in the chick embryo and suggest parameters to electroporate cells within different target tissues that with some modifications may be applicable to a wide range of developmental stages and other embryo model organisms.

摘要

卵内电穿孔可实现非病毒质粒DNA和/或吗啉代寡核苷酸的转染,从而对感兴趣的细胞中的基因进行荧光标记和/或干扰其功能。然而,由于电极的放置和尺寸限制,将电穿孔靶向导入胚胎的特定亚区域可能具有挑战性。在这里,我们描述了鸡胚卵内电穿孔的基本技术,并提出了在不同靶组织内对细胞进行电穿孔的参数,这些参数经过一些修改后可能适用于广泛的发育阶段和其他胚胎模式生物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5ba/6820354/c3de44bdc221/nihms-1056295-f0001.jpg

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