美国斯坦福大学Lei S. Qi研究组报道了活细胞和神经元空间转录组的可编程控制。这一研究成果发表在2025年5月21日出版的国际学术期刊《自然》上。
在这里，研究团队提出了CRISPR介导的转录组组织（CRISPR-TO），这是一种利用RNA引导的核酸酶死亡dCas13可编程控制活细胞内源性RNA定位的系统。CRISPR-TO能够将内源性RNA靶向定位到不同的亚细胞区室，包括各种细胞类型的线粒体外膜、p体、应激颗粒、端粒和核应激体。它允许诱导和可逆的RNA沿着微管通过运动蛋白双向运输，促进实时操作和监测活细胞中的RNA定位动力学。在初级皮层神经元中，课题组发现重新定位的mRNA沿着神经突和神经突尖端进行局部翻译，并与核糖体共同运输，β-肌动蛋白mRNA的定位增强了动态丝状突起的形成，抑制了轴突再生。在初级神经元中启用CRISPR-TO筛选鉴定Stmn2 mRNA定位是神经突生长的驱动因素。通过对空间转录组的大规模扰动，CRISPR-TO弥补了测序和成像技术留下的关键空白，为活细胞和生物体中RNA定位的高通量功能询问提供了一个多功能平台。
据介绍，空间RNA组织在多种细胞过程和疾病中起着关键作用。然而，由于在特定亚细胞区域干扰内源性RNA的技术有限，空间转录组的功能意义在很大程度上仍未被探索。
附：英文原文
Title: Programmable control of spatial transcriptome in live cells and neurons
Author: Han, Mengting, Fu, Maylin L., Zhu, Yanyu, Choi, Alexander A., Li, Emmy, Bezney, Jon, Cai, Sa, Miles, Leann, Ma, Yitong, Qi, Lei S.
Issue&Volume: 2025-05-21
Abstract: Spatial RNA organization has a pivotal role in diverse cellular processes and diseases1,2,3,4. However, functional implications of the spatial transcriptome remain largely unexplored due to limited technologies for perturbing endogenous RNA within specific subcellular regions1,5. Here we present CRISPR-mediated transcriptome organization (CRISPR-TO), a system that harnesses RNA-guided, nuclease-dead dCas13 for programmable control of endogenous RNA localization in live cells. CRISPR-TO enables targeted localization of endogenous RNAs to diverse subcellular compartments, including the outer mitochondrial membrane, p-bodies, stress granules, telomeres and nuclear stress bodies, across various cell types. It allows for inducible and reversible bidirectional RNA transport along microtubules via motor proteins, facilitating real-time manipulation and monitoring of RNA localization dynamics in living cells. In primary cortical neurons, we demonstrate that repositioned mRNAs undergo local translation along neurites and at neurite tips, and co-transport with ribosomes, with β-actin mRNA localization enhancing the formation of dynamic filopodial protrusions and inhibiting axonal regeneration. CRISPR-TO-enabled screening in primary neurons identifies Stmn2 mRNA localization as a driver of neurite outgrowth. By enabling large-scale perturbation of the spatial transcriptome, CRISPR-TO bridges a critical gap left by sequencing and imaging technologies, offering a versatile platform for high-throughput functional interrogation of RNA localization in living cells and organisms.
DOI: 10.1038/s41586-025-09020-z
Source: https://www.nature.com/articles/s41586-025-09020-z