First Knockdown Gene Expression in Bat (Hipposideros armiger) Brain Mediated by Lentivirus
First Knockdown Gene Expression in Bat (Hipposideros armiger) Brain Mediated by Lentivirus
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慢病毒介导的蝙蝠大脑中首次敲低基因表达
DOI：
10.1007/s12033-012-9596-6
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发表时间：
2013-06-01
影响因子：
2.6
通讯作者：
Sun, Yi
中科院分区：
文献类型：
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作者：
Chen, Qi;Zhu, Tengteng;Sun, Yi
慢病毒介导的RNA干扰（RNAi）是研究基因在体外和体内功能的有力实验工具。它具有转基因技术所缺乏的优点。然而，由于缺乏标准的脑图谱和遗传信息，在体内的应用很难应用于非模式生物的中枢神经系统。在此，我们首次报道了一种用于蝙蝠脑组织的体内基因传递系统的开发，该系统基于表达短发夹RNA （shRNA）的慢病毒（LV）载体，靶向希波sideros armigerforkhead box P2 （FoxP2）。用携带编码foxp2shrna的载体体外转染HEK 293T细胞，验证了其敲除效率。假病毒颗粒通过立体定向脑内显微注射进入h的前扣带皮层。骑士的扈从。foxp2是主要的兴趣，因为它在感觉运动协调和可能在回声定位中的作用。随后的原位杂交证实了靶基因的体内沉默。本报告表明，lv介导的RNAi表达可以在蝙蝠这种非模式生物中实现有效的基因沉默，并将有助于阐明蝙蝠基因的功能。
Lentivirus-mediated RNA interference (RNAi) is a potent experimental tool for investigating gene functions in vitro and in vivo. It has advantages that transgenic technology lacks. However, in vivo applications are difficult to apply in the central nervous system of non-model organisms due to the lack of a standard brain atlas and genetic information. Here, we report the development of an in vivo gene delivery system used in bat brain tissue for the first time, based on lentivirus (LV) vectors expressing short hairpin RNA (shRNA) targetingHipposideros armigerforkhead box P2 (FoxP2). In vitro transfection into HEK 293T cell with the vector bearing the cassettes encodingFoxP2shRNA verified the knockdown efficiency. Pseudovirus particles were administered via stereotactic intracerebral microinjection into the anterior cingulate cortex ofH. armiger.FoxP2is of major interest because of its role in sensorimotor coordination and probably in echolocation. Subsequent in situ hybridization validated the in vivo silencing of the target gene. This report demonstrates that LV-mediated expression of RNAi could achieve effective gene silencing in bats, a non-model organism, and will assist in elucidating the functions of bat genes.