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。
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Dev Biol. 2015 Nov 1;407(1):12-25. doi: 10.1016/j.ydbio.2015.08.011. Epub 2015 Aug 13.
2
Parameters affecting efficiency of in ovo electroporation of the avian neural tube and crest.影响禽神经管和嵴卵内电穿孔效率的参数。
Dev Dyn. 2014 Nov;243(11):1440-7. doi: 10.1002/dvdy.24163. Epub 2014 Jul 29.
3
Expression of actin-binding proteins and requirement for actin-depolymerizing factor in chick neural crest cells.鸡神经嵴细胞中肌动蛋白结合蛋白的表达及肌动蛋白解聚因子的需求
Dev Dyn. 2014 May;243(5):730-8. doi: 10.1002/dvdy.24105.
4
Foxi3 is necessary for the induction of the chick otic placode in response to FGF signaling.Foxi3 对于 FGF 信号诱导鸡胚耳嵴的形成是必需的。
Dev Biol. 2014 Jul 15;391(2):158-69. doi: 10.1016/j.ydbio.2014.04.014. Epub 2014 Apr 26.
5
A series of normal stages in the development of the chick embryo.鸡胚胎发育的一系列正常阶段。
J Morphol. 1951 Jan;88(1):49-92.
6
Expression and function of transcription factor cMyb during cranial neural crest development.转录因子cMyb在颅神经嵴发育过程中的表达及功能
Mech Dev. 2014 May;132:38-43. doi: 10.1016/j.mod.2014.01.005. Epub 2014 Feb 6.
7
Neurotrophic factor NT-3 displays a non-canonical cell guidance signaling function for cephalic neural crest cells.神经营养因子 NT-3 对头颅神经嵴细胞表现出非经典的细胞导向信号功能。
Eur J Cell Biol. 2013 Aug-Sep;92(8-9):264-79. doi: 10.1016/j.ejcb.2013.10.006. Epub 2013 Nov 1.
8
Annexin a6 modulates chick cranial neural crest cell emigration.膜联蛋白 A6 调节鸡颅神经嵴细胞的迁移。
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9
Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis.鸡胚电穿孔对内源基因表达的影响：全基因组分析。
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10
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