近日，中国科学院脑科学与智能技术卓越创新中心刘真及其研究团队报道了猕猴脑细胞类型特异性增强子的鉴定与应用。相关论文于2025年7月10日发表于国际顶尖学术期刊《细胞》杂志上。
利用猕猴大脑的单细胞RNA和ATAC测序结合体内筛选，课题组人员发现了一组能够在特定细胞类型中驱动靶向基因表达的增强子。由这些增强子驱动的AAV载体成功靶向层特异性谷氨酸能神经元、GABA能中间神经元亚型、星形胶质细胞和少突胶质细胞，具有高特异性。跨物种比较显示，一些猕猴增强子在物种间是保守的和功能性的，但在猕猴中具有层特异性靶向的增强子在小鼠中没有标记神经元，这突出了皮质CREs的进化差异。使用两个增强子的FLPo依赖交叉方法进一步提高了靶向精度。这些增强-AAVs通过监测和操纵猕猴视觉皮层的活动来验证，为解剖灵长类动物的神经回路功能提供了有价值的工具。
据了解，用于监测和操纵神经元活动的遗传靶向方法在灵长类动物大脑研究中没有广泛的主题，主要是由于缺乏细胞类型特异性靶向方法。
附：英文原文
Title: Identification and application of cell-type-specific enhancers for the macaque brain
Author: Ling Li, Yiming Huang, Dengyu Lu, Jiqiang Fu, Luyan Wu, Zheyuan Chen, Haiou Liao, Jiaqi Zhang, Li Li, Tianyue Gu, Fang He, Hean Liu, Xiangqing Leng, Jing Tang, Jiaqi Yan, Cirong Liu, Chao Li, Huapin Huang, Lina Wang, Longqi Liu, Chengyu Li, Mu-ming Poo, Hao Li, Yidi Sun, Zhen Liu
Issue&Volume: 2025-07-10
Abstract: Genetic targeting methods for monitoring and manipulating neuronal activity are not widely used for studying the primate brain, largely owing to the lack of a cell-type-specific targeting method. Using single-cell RNA and ATAC sequencing of macaque brains combined with in vivo screening, we identified a large set of enhancers capable of driving targeted gene expression in specific cell types. AAV vectors driven by these enhancers successfully targeted layer-specific glutamatergic neurons, GABAergic interneuron subtypes, astrocytes, and oligodendrocytes with high specificity. Cross-species comparison revealed that some macaque enhancers are conserved and functional across species, but enhancers with layer-specific targeting in macaques did not label neurons in mice, highlighting evolutionary differences in cortical CREs. Targeting precision was further improved using a FLPo-dependent intersectional approach with two enhancers. These enhancer-AAVs were validated by monitoring and manipulating activity in macaque visual cortex, providing valuable tools to dissect primate neural circuit functions.
DOI: 10.1016/j.cell.2025.06.040
Source: https://www.cell.com/cell/abstract/S0092-8674(25)00742-1