高分辨率FFPE切片上总RNA的空间定位立体测序V2，这一成果由基因组与多组学技术国家重点实验室徐讯课题组经过不懈努力而取得。该研究于2025年8月28日发表于国际一流学术期刊《细胞》杂志上。
课题组研究人员介绍了Stereo-seq V2，它采用随机引物在FFPE切片上捕获rna并对其进行原位测序，并提供单细胞分辨率。基于随机引物的策略提供了无偏的转录物捕获和统一的基因体覆盖，这增加了对标记基因的敏感性，非聚腺苷化（聚(A)） RNA分析的效率和免疫库覆盖。该团队证明了Stereo-seq V2在三阴性乳腺癌（TNBC）切片的临床FFPE样本上的最佳表现，并确定了肿瘤特异性的选择性剪接事件。在结核分枝杆菌（Mtb）感染的motheme模型中，课题组利用Stereo-seq V2同时监测宿主和病原体转录组的基因表达动态。研究组还组装了免疫库并鉴定了mtb特异性BCR克隆，这些克隆也可以在人类结核肺样本中观察到。这些结果突出了Stereo-seq V2在生物医学研究和个性化医疗方面的潜力。
据了解，对福尔马林固定石蜡包埋（FFPE）样品进行总RNA分析是临床实践中主要的样品保存方法，但对当前的空间转录组学技术仍然具有挑战性。
附：英文原文
Title: Stereo-seq V2: Spatial mapping of total RNA on FFPE sections with high resolution
Author: Yu Zhao, Young Li, Ying He, Junqi Wu, Yi Liu, Xinxing Li, Zhaoxun Li, Qiaomei Yuan, Jialuo Li, Xinya Zhang, Xiaole Hu, Guifang Wu, Ying Qiu, Jie Yuan, Xin Huang, Mengru Xu, Mei Yang, Jing Guo, Tingting Cao, Qiuling Chen, Dandan Li, Jiajun Zhang, Xiaoxi Zhou, Xia Jiang, Fan Zhu, Xuan Dong, Rong Xiang, Hailin Pan, Lei Han, Ziqing Deng, Haohao Deng, Yan Zhang, Maoyuan Liu, Qin Wu, Gaoyang Wang, Jie Zhai, Weihong Tan, Xiawei Liu, Zehao Wang, Shanshan Li, Tianbi Duan, Longqi Liu, Ao Chen, Haiying Liu, Chang Chen, Sha Liao, Xun Xu
Issue&Volume: 2025-08-28
Abstract: Performing total RNA profiling on formalin-fixed, paraffin-embedded (FFPE) samples, the predominant sample conservation method in clinical practice, remains challenging for current spatial transcriptomics techniques. Here, we introduce Stereo-seq V2, which employs random primers to capture and sequence RNAs in situ on FFPE sections and provides single-cell resolution. The random-priming-based strategy offers unbiased transcript capturing and uniform gene body coverage, which increase the sensitivity to marker genes, the efficiency of non-polyadenylation (poly(A)) RNA profiling, and immune repertoire coverage. We demonstrated the robust performance of Stereo-seq V2 on clinical FFPE samples using triple-negative breast cancer (TNBC) sections and identified tumor-specific alternative splicing events. In a Mycobacterium tuberculosis (Mtb)-infected mouse model, we monitored gene expression dynamics of host and pathogen transcriptomes simultaneously by utilizing Stereo-seq V2. We also assembled immune repertoires and identified Mtb-specific BCR clones, which could also be observed in human tuberculous lung samples. These results highlight Stereo-seq V2’s potential in biomedical research and personalized medicine.
DOI: 10.1016/j.cell.2025.08.008
Source: https://www.cell.com/cell/abstract/S0092-8674(25)00922-5