小柯机器人

英国医学研究委员会分子生物学实验室Marco Tripodi和Ernesto Ciabatti共同合作，近期取得重要工作进展。他们研究将长期回路映射和网络转录组学与SiR-N2c相结合。相关研究成果2023年3月2日在线发表于《自然—方法学》杂志上。

据介绍，回路神经科学的一个令人兴奋的前沿是神经网络映射和单细胞基因组学之间的交叉点。单突触狂犬病病毒为回路映射方法与组学方法的结合提供了一个有前景的平台。然而，三个关键限制阻碍了从狂犬病图谱中提取生理学意义的基因表达谱：固有的病毒细胞毒性、高病毒免疫原性和病毒诱导的细胞转录调节改变。这些因素改变了受感染神经元及其邻近细胞的转录和翻译谱。

为了克服这些限制，研究人员对免疫原性较弱的狂犬病毒株CVS-N2c进行了自失活基因组修饰，以产生自失活的CVS-N2c狂犬病病毒（SiR-N2c）。SiR-N2c不仅消除了不期望的细胞毒性效应，而且还显著减少了受感染神经元中的基因表达变化，并抑制了先天性和获得性免疫应答的募集，从而能够使用单细胞基因组方法对神经网络及其遗传特征进行开放式干预。

附：英文原文

Title: Combining long-term circuit mapping and network transcriptomics with SiR-N2c

Author: Lee, Hassal, Ciabatti, Ernesto, Gonzlez-Rueda, Ana, Williams, Elena, Nugent, Fiona, Mookerjee, Souradip, Morgese, Fabio, Tripodi, Marco

Issue&Volume: 2023-03-02

Abstract: An exciting frontier in circuit neuroscience lies at the intersection between neural network mapping and single-cell genomics. Monosynaptic rabies viruses provide a promising platform for the merger of circuit mapping methods with -omics approaches. However, three key limitations have hindered the extraction of physiologically meaningful gene expression profiles from rabies-mapped circuits: inherent viral cytotoxicity, high viral immunogenicity and virus-induced alteration of cellular transcriptional regulation. These factors alter the transcriptional and translational profiles of infected neurons and their neighboring cells. To overcome these limitations we applied a self-inactivating genomic modification to the less immunogenic rabies strain, CVS-N2c, to generate a self-inactivating CVS-N2c rabies virus (SiR-N2c). SiR-N2c not only eliminates undesired cytotoxic effects but also substantially reduces gene expression alterations in infected neurons and dampens the recruitment of innate and acquired immune responses, thus enabling open-ended interventions on neural networks and their genetic characterization using single-cell genomic approaches.

DOI: 10.1038/s41592-023-01787-1

Source: https://www.nature.com/articles/s41592-023-01787-1

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex