Comparison of the efficacy of five adeno-associated virus vectors for transducing dorsal raphé nucleus cells in the mouse.
Comparison of the efficacy of five adeno-associated virus vectors for transducing dorsal raphé nucleus cells in the mouse.
复制标题
比较五种腺相关病毒载体转导小鼠中缝背核细胞的功效。
DOI：
10.1016/j.jneumeth.2014.07.005
复制
发表时间：
2014
影响因子：
3
通讯作者：
Jacobson,Lauren
中科院分区：
文献类型：
--
作者：
Vincent,Melanie;Gao,Guangping;Jacobson,Lauren
背景用腺相关病毒(AAV)的2型血清可以将基因输送到不同的大脑区域。伪型AAV2载体由包装在另一种血清型衣壳中的AAV2基因组组成，提高了病毒转导的效率。伪型AAV2载体的转导依赖于细胞类型、脑区域和发育阶段。中缝背核(DRN)和中缝正中核为前脑区域提供了大部分的5-羟色胺，并与抑郁症和焦虑症的病理和治疗有关。病毒载体技术与小鼠立体定向手术相结合，提供了一种区别DRN和中缝中核基因功能的方法。新方法由于AAV2对DRN的转导效率尚未确定，我们在C57BL/6J背景下测试了AAV2伪型是否比标准血清型(AAV2/2)更有效地转导成年雄性小鼠的DRN细胞。结果尽管在注射后15天，不同载体之间的转导没有显著差异，但在注射后30天，伪型AAV2/9和AAV2/rh.10载体对DRN的转导显著高于AAV2/2和AAV2/1载体。假型AAV2/1和AAV2/5转导DRN细胞的效率虽然不显著高于AAV2/2。与现有方法相比，在相同滴度下，所有受试的假型AAV在注射后30天都比标准AAV2/2血清型更有效地转导DRN。结论我们的结果支持使用伪型AAV2/9和AAV2/r.10研究DRN中的基因缺失或过表达。
BackgroundDelivery of genes to various brain regions can be accomplished using serotype 2 of the adeno-associated virus (AAV). Pseudotype AAV2 vectors, composed of the AAV2 genome packaged in the capsid of an alternative serotype, have increased efficiency of viral transduction. Transduction of pseudotype AAV2 vectors depends on cell type, brain region and stage of development. The dorsal raphé nucleus (DRN) and median raphé provides the majority of serotonin to forebrain regions and are implicated in the pathology and treatment of depression and anxiety. Viral vector technology in combination with stereotaxic surgery in mice provides a means to differentiate gene function in the DRN compared to the median raphé nucleus.New methodSince AAV transduction efficiency has not yet been characterized for the DRN, we tested if AAV2 pseudotypes are more efficient than a standard serotype (AAV2/2) in transducing DRN cells in adult male mice on a C57BL/6J background.ResultsAlthough transduction did not differ significantly among vectors by 15 days post-injection, pseudotype AAV2/9 and AAV2/rh.10 vectors achieved significantly greater transduction of the DRN than did AAV2/2 and AAV2/1 vectors by 30 days post-injection. Pseudotypes AAV2/1 and AAV2/5 tended, although not significantly, to transduce DRN cells more efficiently than did AAV2/2.Comparison with existing methodsAt the same titer, all pseudotype AAV tested tended to transduce the DRN more efficiently than standard AAV2/2 serotype at 30 days post-injection.ConclusionsOur results support the use of pseudotype AAV2/9 and AAV2/rh.10 for studying gene deletion or overexpression in the DRN.