Name: gene transfer into older chicken embryos by ex ovo electroporation
Uploaded: 2012-07-27T12:00:00
Description: Watch this Scientific Journal Video about Gene Transfer into Older Chicken Embryos by ex ovo Electroporation at JoVE.com

这项工作是由德国研究基金会（DFG; LU1455/1-1）的资助支持。

1。 前大毛文化<ol><li>在强制通风孵化器（BSS160，Ehret，德国）在37.5°C间新鲜受精卵奠定孵化湿度60％到他们的长边。在12°C储存时间超过一个星期的鸡蛋不应使用。 </li><li> 2.5天（约阶段HH17）孵化后，取出鸡蛋和标签顶端，用铅笔表明开裂方向。 </li><li>在打击之前，倒入一个干净的大培养皿（直径145毫米;格雷纳生物公司之一，德国），约20毫升无菌蒸馏水。 </li><li>到大培养皿中，将另一...

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	<li>Momose, T., Tonegawa, A., Takeuchi, J., Ogawa, H., Umesono, K., Yasuda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+targeting+of+gene+expression+in+chick+embryos+by+microelectroporation.">Efficient targeting of gene expression in chick embryos by microelectroporation.</a> Dev. Growth Differ. 41, 335-344 (1999).</li><li>Luo, J., Ju, M. J., Lin, J., Yan, X., Markus, A., Mix, E., Rolfs, A., Redies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cadherin-20+expression+by+motor+neurons+is+regulated+by+Sonic+hedgehog+during+spinal+cord+development.">Cadherin-20 expression by motor neurons is regulated by Sonic hedgehog during spinal cord development.</a> NeuroReport. 20, 365-370 (2009).</li><li>Luo, J., Ju, M. J., Redies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regionalized+cadherin-7+expression+by+radial+glia+is+regulated+by+Shh+and+Pax7+during+chicken+spinal+cord+development.">Regionalized cadherin-7 expression by radial glia is regulated by Shh and Pax7 during chicken spinal cord development.</a> Neuroscience. 142, 1133-1143 (2006).</li><li>Nakamura, H., Katahira, T., Sato, T., Watanabe, Y., Funahashi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gain-+and+loss-of-function+in+chick+embryos+by+electroporation.">Gain- and loss-of-function in chick embryos by electroporation.</a> Mech. Dev. 121, 1137-1143 (2004).</li><li>Dai, F., Yusuf, F., Farjah, G. H., Brand-Saberi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNAi-induced+targeted+silencing+of+developmental+control+genes+during+chicken+embryogenesis.">RNAi-induced targeted silencing of developmental control genes during chicken embryogenesis.</a> Dev. Biol. 258, 80-90 (2005).</li><li>Rao, M., Baraban, J. H., Rajaii, F., Sockanathan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+comparative+study+of+RNAi+methodologies+by+in+ovo+electroporation+in+the+chick+embryo.">In vivo comparative study of RNAi methodologies by in ovo electroporation in the chick embryo.</a> Dev. Dyn. 231, 592-600 (2004).</li><li>Kos, R., Tucker, R. P., Hall, R., Duong, T. D., Erickson, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+introducing+morpholinos+into+the+chicken+embryo.">Methods for introducing morpholinos into the chicken embryo.</a> Dev. Dyn. 226, 470-477 (2003).</li><li>Hamburger, V., Hamilton, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+series+of+normal+stages+in+the+development+of+the+chick+embryo.">A series of normal stages in the development of the chick embryo.</a> J. Morphol. 88, 49-92 (1951).</li><li>Auerbach, R., Kubai, L., Knighton, D., Folkman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+procedure+for+the+long-term+cultivation+of+chicken+embryos.">A simple procedure for the long-term cultivation of chicken embryos.</a> Dev. Biol. 10, 391-394 (1974).</li><li>Luo, J., Treubert-Zimmermann, U., Redies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cadherins+guide+migrating+Purkinje+cells+to+specific+parasagittal+domains+during+cerebellar+development.">Cadherins guide migrating Purkinje cells to specific parasagittal domains during cerebellar development.</a> Mol. Cell. Neurosci. 25, 138-152 (2004).</li><li>Luo, J., Redies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+ovo+electroporation+for+gene+transfer+into+older+chicken+embryos.">Ex ovo electroporation for gene transfer into older chicken embryos.</a> Dev. Dyn. 233, 1470-1477 (2005).</li><li>Treubert-Zimmermann, U., Heyers, D., Redies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+axons+to+specific+fiber+tracts+in+vivo+by+altering+cadherin+expression.">Targeting axons to specific fiber tracts in vivo by altering cadherin expression.</a> J. Neurosci. 22, 7617-7626 (2002).</li></ol>

该协议提供了旧鸡胚前OVO电（例如，将在E6顶盖）的转基因指南。这种方法延长使用电老鸡胚胎，并在稍后阶段研究基因在体内的功能可以方便地应用于许多实验室。至少从E4类E7的胚胎可以用这种方法成功的电穿孔和电穿孔胚胎可以存活直到E15 11。除了 ​​大脑，这种方法还可以用于基因转移成鸡肢体制度11。 应特别注意支付给由前OVO电?...

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> electroporator </td><td>日本NEPA基因， </td><td> CUY21的编辑</td></tr><tr><td>电极</td><td>日本NEPA基因， </td><td> CUY661-3x7 </td></tr><tr><td>鸡蛋孵化器</td><td> ehret，德国</td><td> BSS160 </td></tr><tr><td>胚胎孵化器</td><td>粘合剂有限公司，德国</td><td></td></tr><tr><td>荧光显微镜</td><td> KEYENCE，德国公司</td><td> BZ-8000 </td></tr><tr><td>培养皿</td><td>格雷纳生物，其中有限责任公司，德国</td><td></td></tr><tr><td>微电极拔</td><td>世界精密仪器，德国</td><td>的Pul-100 </td></tr></tbody></table>

gene transfer into older chicken embryos by ex ovo electroporation

鸡胚胎为研究胚胎发育过程中基因功能和调控提供了一个很好的模型系统。 在OVO电是一个强大的过度表达外源基因的方法或下调体内的内源性基因在鸡胚1。不同的结构，如DNA质粒编码基因的小干扰RNA（siRNA）的质粒5，小合成的RNA寡核苷酸6，和吗啉反义寡核苷酸7 2-4，可以很容易地通过电转染鸡胚胎。然而， 在OVO电的应用是有限的胚胎早期孵化阶段（年龄小于阶段HH20 -据汉堡和汉密尔顿）8，有一些缺点是其在胚胎中的应用，在稍后阶段（比HH22阶段老年人-约3.5发展天）。例如，在稍后阶段卵黄膜是通常盟友坚持应当膜，并在外壳打开一个窗口，导致血管破裂，导致胚胎死亡;年纪较大的胚胎卵黄囊和尿囊血管，它是很难访问和操纵胚胎覆盖旧胚胎移动大力通过在shell中比较小的窗口，是难以控制的方向。 在这个协议中，我们证明前OVO电穿孔法基因转移到鸡胚胎后期（比HH22阶段旧）。对于前大毛电，是胚胎培养在培养皿9和卵黄和尿囊血管被广泛流传。在这些条件下，年龄较大的鸡胚胎很容易访问和操纵。因此，这种方法克服缺点，适用于年龄较大的鸡胚胎的OVO电。质粒使用这种方法，可以很容易地转成不同的部分老鸡胚胎10-12。

Watch this Scientific Journal Video about Gene Transfer into Older Chicken Embryos by ex ovo Electroporation at JoVE.com

Gene Transfer into Older Chicken Embryos by ex ovo Electroporation

介绍将在以后的孵化阶段（比汉堡和汉密尔顿阶段（HH）22岁以上）的鸡胚胎的基因转移方法。这种方法克服缺点<em&gt;在OVO</em&gt;电适用于年龄较大的鸡胚胎是一种有用的技术，研究基因功能和调控，在旧的发展阶段。

基因转移到老年人的鸡胚<em&gt;前OVO</em&gt;电穿孔

The chicken embryo provides an  excellent model system for studying gene function and regulation during  embryonic development. In ovo electroporation is ...

Research

JoVE Journal

Biology

Gene Transfer into Older Chicken Embryos by ex ovo Electroporation

Nuclear Transfer into Mouse Oocytes

小鼠卵母细胞的细胞核移植到

Nuclear transfer into an unfertilized oocyte can restore developmental potential to a differentiated cell. This demonstrates that the processes underlying ...

科研

生物学

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos

E15小鼠胚胎在子宫内侧脑室注射和电穿孔

In-utero in-vivo injection and electroporation of the embryonic mouse neocortex provides a powerful tool for the manipulation of individual progenitors ...

Placing Growth Factor-Coated Beads on Early Stage Chicken Embryos

配售早期鸡胚生长因子包被的磁珠

The neural tube expresses many proteins in specific spatiotemporal patterns during development.   These proteins have been shown to be critical for cell ...

In Ovo Electroporations of HH Stage 10 Chicken Embryos

在OVO的HH阶段10个鸡胚Electroporations

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology.  With the advent of ...

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

主电穿孔的小鼠骨髓造血细胞培养制备

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

In utero and ex vivo Electroporation for Gene Expression in Mouse Retinal Ganglion Cells

In utero and ex vivo Electroporation for Gene Expression in Mouse Retinal Ganglion Cells

在子宫内和前体内的基因表达在小鼠视网膜神经节细胞的电穿孔

The retina and its sole output neuron, the retinal ganglion cell (RGC), comprise an excellent model in which to examine biological questions such as cell ...

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

小鸡前OVO文化前OVO CAM含量：它是如何真正起作用

Chicken eggs in the early phase of breeding are between in vitro and in vivo systems and provide a vascular test environment not only to study ...

Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

使用基因脉冲MXcell电穿孔系统主细胞的转染效率高

Xenotransplantation of Human Stem Cells into the Chicken Embryo

人类干细胞异种移植到鸡胚

The chicken embryo is a classical animal model for studying normal embryonic and fetal development and for xenotransplantation experiments to study the ...

An ex-ovo Chicken Embryo Culture System Suitable for Imaging and Microsurgery Applications

An ex-ovo Chicken Embryo Culture System Suitable for Imaging and Microsurgery Applications

一个 EX - OVO鸡胚胎培养系统适用于成像和显微外科的应用

Understanding the relationships between genetic and microenvironmental factors that drive normal and malformed embryonic development is fundamental for ...