Cell-to-cell and genome-to-genome variability of adenovirus transcription tuned by the cell cycle

• DOI: 10.1242/jcs.252544
• Publication Date: 2020-09-11T19:05:13Z
• Authors: Maarit Suomalainen; Vibhu Prasad; Abhilash Kannan; Urs F. Greber
• Keywords: Adenovirus genome; Transcription

In clonal cultures, not all cells are equally susceptible to virus infection, and the mechanisms underlying this poorly understood. Here, we developed image-based single-cell measurements scrutinize heterogeneity of adenovirus (AdV) infection. AdV delivers, transcribes replicates a linear double-stranded DNA genome in nucleus. We measured abundance viral transcripts using single-molecule RNA fluorescence situ hybridization (FISH) incoming 5-ethynyl-2'-deoxycytidine (EdC)-tagged genomes copper(I)-catalyzed azide-alkyne cycloaddition (click) reaction. Surprisingly, expression immediate early gene E1A only moderately correlated with number cell Intranuclear genome-to-genome was found at level transcription and, accordance, individual exhibited heterogeneous replication activity. By analyzing cycle state, that G1 highest displayed increased correlation between copy numbers. The combined procedures described here ideally suited explore cell-to-cell variability range different settings, including innate immune response.