近日，美国加州理工学院教授Viviana Gradinaru研究团队报道了AAV载体的空间基因组学揭示了转录串扰机制，使大的遗传货物靶向递送。这一研究成果于2025年3月20日发表在国际顶尖学术期刊《自然—生物技术》上。

在这项研究中，研究团队以空间基因组学为主题，表明单个AAV基因组之间的转录串扰通过将远端作用的调控元件分离到第二个AAV基因组中，为大货物的细胞类型特异性表达提供了一种通用方法。该研究组鉴定并分析了携带11种不同增强子的AAV基因组的转录串扰。该课题组开发了空间基因组学方法来鉴定和定位AAV基因组及其在培养细胞和组织中的串联形式，该课题组在这里证明了转录串扰依赖于串联结构的形成。最后，研究团队利用转录串扰以系统管理的工程AAV以细胞类型特异性的方式驱动3.2-kb Cas9货物的表达，并且研究团队在概括已知疾病表型的野生型小鼠中展示了AAV传递的微创细胞类型特异性基因编辑。

研究人员表示，细胞类型特异性调控元件（如增强子）可以将重组腺相关病毒主题（AAV）直接表达到特定的细胞类型，但这种方法受到AAV相对较小的包装容量的限制。

附：英文原文

Title: Spatial genomics of AAV vectors reveals mechanism of transcriptional crosstalk that enables targeted delivery of large genetic cargo

Author: Coughlin, Gerard M., Borsos, Mt, Barcelona, BreAnna H., Appling, Nathan, Mayfield, Acacia M. H., Mackey, Elisha D., Eser, Rana A., Jackson, Cameron R., Chen, Xinhong, Kumar, Sripriya Ravindra, Gradinaru, Viviana

Issue&Volume: 2025-03-20

Abstract: Cell-type-specific regulatory elements such as enhancers can direct expression of recombinant adeno-associated viruses (AAVs) to specific cell types, but this approach is limited by the relatively small packaging capacity of AAVs. In this study, we used spatial genomics to show that transcriptional crosstalk between individual AAV genomes provides a general method for cell-type-specific expression of large cargo by separating distally acting regulatory elements into a second AAV genome. We identified and profiled transcriptional crosstalk in AAV genomes carrying 11 different enhancers active in mouse brain. We developed spatial genomics methods to identify and localize AAV genomes and their concatemeric forms in cultured cells and in tissue, and we demonstrate here that transcriptional crosstalk is dependent upon concatemer formation. Finally, we leveraged transcriptional crosstalk to drive expression of a 3.2-kb Cas9 cargo in a cell-type-specific manner with systemically administered engineered AAVs, and we demonstrate AAV-delivered, minimally invasive, cell-type-specific gene editing in wild-type mice that recapitulates known disease phenotypes.

DOI: 10.1038/s41587-025-02565-4

Source: https://www.nature.com/articles/s41587-025-02565-4