ABSTRACT
Viral production and transduction have seen a considerable increase in use for gene therapies and especially vaccines (e.g., COVID-19) due to their abilities to deliver a significant amount of genetic information and integrate it into the genome. However, challenges associated with viral production/transduction involve steps that are very time consuming, manually intensive, and laborious. In efforts to expedite this process, we have created a microfluidic methodology that will provide a “hands-off” workflow in the genome engineering pipeline. In this work, we developed a platform which can generate lentiviral particles on-demand containing the gene-editing machinery that will be able to modify target breast cancer cell lines. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.
ABSTRACT
A key to tackling the coronavirus disease 2019 (COVID-19) pandemic is to understand how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) manages to outsmart host antiviral defense mechanisms. Stress granules (SGs), which are assembled during viral infection and function to sequester host and viral mRNAs and proteins, are part of the antiviral responses. Here, we show that the SARS-CoV-2 nucleocapsid (N) protein, an RNA binding protein essential for viral production, interacted with Ras-GTPase-activating protein SH3-domain-binding protein (G3BP) and disrupted SG assembly, both of which require intrinsically disordered region1 (IDR1) in N protein. The N protein partitioned into SGs through liquid-liquid phase separation with G3BP, and blocked the interaction of G3BP1 with other SG-related proteins. Moreover, the N protein domains important for phase separation with G3BP and SG disassembly were required for SARS-CoV-2 viral production. We propose that N protein-mediated SG disassembly is crucial for SARS-CoV-2 production.