Use of lentiviral vectors to deliver and express bicistronic transgenes in developing chicken embryos.
Use of lentiviral vectors to deliver and express bicistronic transgenes in developing chicken embryos.
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DOI：
10.1016/j.ymeth.2013.06.026
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发表时间：
2014-04-01
期刊：
影响因子：
4.8
通讯作者：
Berry, Jonathan
中科院分区：
文献类型：
--
作者：
Semple-Rowland, Susan L.;Berry, Jonathan
慢病毒载体携带大的转基因（~ 8 Kb）并有效感染这些基因并将其整合到分裂和非分裂细胞的基因组中的能力使其成为将遗传物质转运到细胞和组织中的理想候选者。考虑到这些载体的性质，它们在发育中的组织，特别是发育中的神经系统的研究中的应用有限，这有点令人惊讶。在过去的几年中，我们利用这些载体的大容量来探索几种双启动子和2A肽双顺反子转基因在发育中的鸡神经视网膜中的表达特性，目的是鉴定在感染细胞中可靠表达多种蛋白质的转基因设计。在这里，我们总结了几个这些转基因在神经视网膜的活动，并提供了详细的方法包装慢病毒和提供病毒进入发育中的鸡胚在卵神经管，程序已在我们的实验室使用了几年的过程中进行了优化。并对注射胚胎的孵化条件进行了讨论。鸡特异性技术将是使用禽类胚胎的研究者最感兴趣的，在感染细胞中可靠表达多种蛋白质的慢病毒载体的开发和包装应该是所有实验需要在发育细胞和组织中操纵和表达多种蛋白质的研究者感兴趣的。
The abilities of lentiviral vectors to carry large transgenes (~ 8Kb) and to efficiently infect and integrate these genes into the genomes of both dividing and non-dividing cells make them ideal candidates for transport of genetic material into cells and tissues. Given the properties of these vectors, it is somewhat surprising that they have seen only limited use in studies of developing tissues and in particular of the developing nervous system. Over the past several years, we have taken advantage of the large capacity of these vectors to explore the expression characteristics of several dual promoter and 2A peptide bicistronic transgenes in developing chick neural retina, with the goal of identifying transgene designs that reliably express multiple proteins in infected cells. Here we summarize the activities of several of these transgenes in neural retina and provide detailed methodologies for packaging lentivirus and delivering the virus into the developing neural tubes of chicken embryos in ovo, procedures that have been optimized over the course of several years of use in our laboratory. Conditions to hatch injected embryos are also discussed. The chicken-specific techniques will be of highest interest to investigators using avian embryos, development and packaging of lentiviral vectors that reliably express multiple proteins in infected cells should be of interest to all investigators whose experiments demand manipulation and expression of multiple proteins in developing cells and tissues.
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影响因子：
4.4
作者：
Greenberg, Kenneth P.;Geller, Scott F.;Flannery, John G.
通讯作者：
Flannery, John G.
影响因子：
3
作者：
Chen F;LoTurco J
通讯作者：
LoTurco J
RCAS-RNAI：一种发育中的鸡视网膜的功能丧失方法。
影响因子：
--
作者：
Harpavat, S;Cepko, CL
通讯作者：
Cepko, CL
DOI：
10.1073/pnas.0610155104
发表时间：
2007-01-16
影响因子：
11.1
作者：
Matsuda, Takahiko;Cepko, Constance L.
通讯作者：
Cepko, Constance L.
影响因子：
46.9
作者：
Fang, JM;Qian, JJ;Jooss, K
通讯作者：
Jooss, K