使用RNA荧光原位杂交-流式细胞术对病毒感染细胞进行定量分析。

Quantification of virus-infected cells using RNA FISH-Flow.

作者信息

Warren Cody J, Barbachano-Guerrero Arturo, Huey Devra, Yang Qing, Worden-Sapper Emma R, Kuhn Jens H, Sawyer Sara L

机构信息

Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210, USA.

BioFrontiers Institute, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80303, USA.

出版信息

STAR Protoc. 2023 May 18;4(2):102291. doi: 10.1016/j.xpro.2023.102291.

DOI:10.1016/j.xpro.2023.102291

PMID:37209094

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10209735/

Abstract

We present a protocol to detect cells that have been infected by RNA viruses. The method, RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow), uses 48 fluorescently labeled DNA probes that hybridize in tandem to viral RNA. RNA FISH-Flow probes can be synthesized to match any RNA virus genome, in either sense or anti-sense, enabling detection of genomes or replication intermediates within cells. Flow cytometry enables high-throughput analysis of infection dynamics within a population at the single cell level. For complete details on the use and execution of this protocol, please refer to Warren et al. (2022)..

摘要

我们提出了一种检测已被RNA病毒感染的细胞的方案。该方法，即RNA荧光原位杂交流式细胞术（RNA FISH-Flow），使用48种荧光标记的DNA探针，这些探针串联杂交到病毒RNA上。RNA FISH-Flow探针可以合成以匹配任何RNA病毒基因组，无论是正义链还是反义链，从而能够检测细胞内的基因组或复制中间体。流式细胞术能够在单细胞水平上对群体内的感染动态进行高通量分析。有关此方案的使用和执行的完整详细信息，请参考沃伦等人（2022年）的文献。