AUTHOR=Kim Hyun-Bum , Brosseau Quentin , Radzio Julia , Wang Jinhui , Muramatsu Hiromi , Kuang Da , Grady M. Sean , Chen H. Isaac , Wolf John A. , Ulyanova Alexandra V. , Bartfai Tamas , Kim Junhyong , Pardi Norbert , Sul Jai-Yoon , Arratia Paulo , Eberwine James TITLE=Single cell phototransfection of mRNAs encoding SARS-CoV2 spike and nucleocapsid into human astrocytes results in RNA dependent translation interference JOURNAL=Frontiers in Drug Delivery VOLUME=4 YEAR=2024 URL=https://www.frontiersin.org/journals/drug-delivery/articles/10.3389/fddev.2024.1359700 DOI=10.3389/fddev.2024.1359700 ISSN=2674-0850 ABSTRACT=

Multi-RNA co-transfection is starting to be employed to stimulate immune responses to SARS-CoV-2 viral infection. While there are good reasons to utilize such an approach, there is little background on whether there are synergistic RNA-dependent cellular effects. To address this issue, we use transcriptome-induced phenotype remodeling (TIPeR) via phototransfection to assess whether mRNAs encoding the Spike and Nucleocapsid proteins of SARS-CoV-2 virus into single human astrocytes (an endogenous human cell host for the virus) and mouse 3T3 cells (often used in high-throughput therapeutic screens) synergistically impact host cell biologies. An RNA concentration-dependent expression was observed where an increase of RNA by less than 2-fold results in reduced expression of each individual RNAs. Further, a dominant inhibitory effect of Nucleocapsid RNA upon Spike RNA translation was detected that is distinct from codon-mediated epistasis. Knowledge of the cellular consequences of multi-RNA transfection will aid in selecting RNA concentrations that will maximize antigen presentation on host cell surface with the goal of eliciting a robust immune response. Further, application of this single cell stoichiometrically tunable RNA functional genomics approach to the study of SARS-CoV-2 biology promises to provide details of the cellular sequalae that arise upon infection in anticipation of providing novel targets for inhibition of viral replication and propagation for therapeutic intervention.