An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector
An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector
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DOI：
10.1186/1471-2202-11-81
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发表时间：
2010-07-13
期刊：
影响因子：
2.4
通讯作者：
Adan, Roger A. H.
中科院分区：
文献类型：
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作者：
de Backer, Marijke W. A.;Fitzsimons, Carlos P.;Adan, Roger A. H.
背景：本研究比较了腺相关病毒（AAV）载体（用 AAV1 衣壳假型化并编码绿色荧光蛋白（GFP））与慢病毒（LV）载体（用 VSV-G 包膜假型化并编码盘状体红色荧光蛋白（dsRed））的转导效率，以研究哪种病毒载体更有效地转导外侧下丘脑或杏仁核。将 LV-dsRed 和 AAV1-GFP 载体混合并注射到成年大鼠的外侧下丘脑或杏仁核中。注射的滴度为 1 x 10(8) 或 1 x 10(9) 基因组拷贝的 AAV1-GFP 和 1 x 10(5) 转导单位的 LV-dsRed。 结果：GFP 和 dsRed 的免疫染色显示，在外侧下丘脑和杏仁核中，AAV1-GFP 转导的细胞明显多于 LV-dsRed。此外，注射的 LV 颗粒数量不容易增加，而 AAV1 颗粒的数量可以轻松增加 100 至 1000 倍。这两种病毒载体似乎都主要转导神经元。结论：这项研究表明，AAV1 载体是在成年大鼠外侧下丘脑和杏仁核中过表达或敲低基因的更好工具，因为 AAV1 比 LV 载体可以转导更多的细胞，而且滴度也更高。 AAV1 载体可以轻松增加以转导感兴趣的区域。
Background: This study compared the transduction efficiencies of an adeno-associated viral (AAV) vector, which was pseudotyped with an AAV1 capsid and encoded the green fluorescent protein (GFP), with a lentiviral (LV) vector, which was pseudotyped with a VSV-G envelop and encoded the discosoma red fluorescent protein (dsRed), to investigate which viral vector transduced the lateral hypothalamus or the amygdala more efficiently. The LV-dsRed and AAV1-GFP vector were mixed and injected into the lateral hypothalamus or into the amygdala of adult rats. The titers that were injected were 1 x 10(8) or 1 x 10(9) genomic copies of AAV1-GFP and 1 x 10(5) transducing units of LV-dsRed.Results: Immunostaining for GFP and dsRed showed that AAV1-GFP transduced significantly more cells than LV-dsRed in both the lateral hypothalamus and the amygdala. In addition, the number of LV particles that were injected can not easily be increased, while the number of AAV1 particles can be increased easily with a factor 100 to 1000. Both viral vectors appear to predominantly transduce neurons.Conclusions: This study showed that AAV1 vectors are better tools to overexpress or knockdown genes in the lateral hypothalamus and amygdala of adult rats, since more cells can be transduced with AAV1 than with LV vectors and the titer of AAV1 vectors can easily be increased to transduce the area of interest.