新南威尔士大学Irina Voineagu团队的一项最新研究显示，在培养的人类星形胶质细胞中进行CRISPRi筛选，发现控制阿尔茨海默病失调基因的远端增强子。相关论文发表在2025年12月18日出版的《自然—神经科学》杂志上。

在这里，该课题组研究人员提出了AstroREG，这是一种通过结合CRISPR抑制（CRISPRi）、单细胞RNA-seq和机器学习产生的人类原代星形胶质细胞中增强基因相互作用的抗性。通过对近1000个PsychENCODE增强子进行功能测试，该团队确定了150多个调节相互作用，揭示了控制关键星形胶质细胞功能和阿尔茨海默病相关基因的增强子。

CRISPRi筛选还为增强子活性的训练和基准预测模型提供了有价值的原代细胞类型的基本事实数据。他们开发了EGrf，一种基于这些数据训练的随机森林（RF）模型，并将其应用于全基因组范围内，以高特异性预测调控相互作用。总之，他们的数据提供了一种关键神经胶质细胞类型增强剂介导调节的全面功能图谱，揭示了大脑功能和疾病。

据悉，与复杂性状相关的遗传变异通常存在于远端增强子中。虽然候选增强子已经在全基因组范围内定位，但它们在特定细胞类型中的功能状态和基因靶点仍不清楚。

附：英文原文

Title: CRISPRi screening in cultured human astrocytes uncovers distal enhancers controlling genes dysregulated in Alzheimer’s disease

Author: Green, Nicole F. O., Sutton, Gavin J., Prez-Burillo, Javier, Wang, Juli, Bagot, Samuel, Danon, Hannah G., Walsh, Kieran, Gokool, Akira, Miles, Samantha A., Yang, Guang, Herring, Charles A., Liang, Yuheng, Pfundstein, Grant, Sytnyk, Vladimir, Alinejad-Rokny, Hamid, Lister, Ryan, Rosenbluh, Joseph, Gagnon-Bartsch, Johann A., Voineagu, Irina

Issue&Volume: 2025-12-18

Abstract: Genetic variants associated with complex traits often lie in distal enhancers. While candidate enhancers have been mapped genome wide, their functional state and gene targets in specific cell types remain unclear. Here we present AstroREG, a resource of enhancer–gene interactions in human primary astrocytes, generated by combining CRISPR inhibition (CRISPRi), single-cell RNA-seq and machine learning. By functionally testing nearly 1,000 PsychENCODE enhancers, we identified more than 150 regulatory interactions, revealing enhancers that control key astrocyte functions and genes implicated in Alzheimer’s disease. The CRISPRi screen also provided valuable ground-truth data from a primary cell type for training and benchmarking prediction models of enhancer activity. We thus developed EGrf, a random forest (RF) model trained on these data, and applied it genome wide to predict regulatory interactions with high specificity. Together, our data provide a comprehensive functional map of enhancer-mediated regulation in a key glial cell type, shedding light on brain function and disease.

DOI: 10.1038/s41593-025-02154-3

Source: https://www.nature.com/articles/s41593-025-02154-3