Suppr超能文献

小鼠下丘和小脑中六种腺相关病毒载体血清型的比较分析。

Comparative analysis of six adeno-associated viral vector serotypes in mouse inferior colliculus and cerebellum.

作者信息

Witteveen Isabelle, Balmer Timothy

机构信息

School of Life Sciences, Arizona State University, Tempe, AZ, 85287, USA.

出版信息

bioRxiv. 2024 Oct 21:2024.10.17.618966. doi: 10.1101/2024.10.17.618966.

Abstract

Adeno-associated viral vector (AAV) serotypes vary in how effectively they express genes across different cell types and brain regions. Here we report a systematic comparison of the AAV serotypes 1, 2, 5, 8, 9, and the directed evolution derived AAVrg, in the inferior colliculus and cerebellum. The AAVs were identical apart from their different serotypes, each having a synapsin promotor and expressing GFP (AAV-hSyn-GFP). Identical titers and volumes were injected into the inferior colliculus and cerebellum of adult male and female mice and brains were sectioned and imaged 2 weeks later. Transduction efficacy, anterograde labeling of axonal projections, and retrograde labeling of somata, were characterized and compared across serotypes. Cell-type tropism was assessed by analyzing the morphology of the GFP-labeled neurons in the cerebellar cortex. In both the cerebellum and inferior colliculus, AAV1 expressed GFP in more cells, labeled a larger volume, and produced significantly brighter labeling than all other serotypes, indicating superior transgene expression. AAV1 labeled more Purkinje cells, unipolar brush cells, and molecular layer interneurons than the other serotypes, while AAV2 labeled a greater number of granule cells. These results provide guidelines for the use of AAVs as gene delivery tools in these regions.

摘要

腺相关病毒(AAV)血清型在不同细胞类型和脑区中表达基因的效率有所不同。在此,我们报告了对AAV血清型1、2、5、8、9以及通过定向进化获得的AAVrg,在下丘和小脑中进行的系统比较。除血清型不同外,这些AAV均相同,每种都具有突触素启动子并表达绿色荧光蛋白(AAV-hSyn-GFP)。将相同滴度和体积的病毒注射到成年雄性和雌性小鼠的下丘和小脑中,2周后对大脑进行切片和成像。对各血清型的转导效率、轴突投射的顺行标记和胞体的逆行标记进行了表征和比较。通过分析小脑皮质中绿色荧光蛋白标记神经元的形态来评估细胞类型嗜性。在下丘和小脑中,AAV1在更多细胞中表达绿色荧光蛋白,标记的体积更大,且标记亮度明显高于所有其他血清型,表明其转基因表达能力更强。与其他血清型相比,AAV1标记的浦肯野细胞、单极刷状细胞和分子层中间神经元更多,而AAV2标记的颗粒细胞数量更多。这些结果为在这些区域使用AAV作为基因传递工具提供了指导。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e173/11526941/2cedc1d7fde0/nihpp-2024.10.17.618966v1-f0001.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验