Broad Institute, Cambridge, MA 02142, USA.
Department of Pharmaceutical Chemistry, Cardiovascular Research Institute, UC San Francisco, San Francisco, CA 94158, USA.
Viruses. 2023 Sep 27;15(10):2005. doi: 10.3390/v15102005.
The SARS-CoV-2 coronavirus has caused worldwide disruption through the COVID-19 pandemic, providing a sobering reminder of the profound impact viruses can have on human well-being. Understanding virus life cycles and interactions with host cells lays the groundwork for exploring therapeutic strategies against virus-related diseases. Fluorescence microscopy plays a vital role in virus imaging, offering high spatiotemporal resolution, sensitivity, and spectroscopic versatility. In this opinion piece, we first highlight two recent techniques, SunTag and StayGold, for the in situ imaging of viral RNA translation and viral assembly. Next, we discuss a new class of genetically encoded fluorogenic protease reporters, such as FlipGFP, which can be customized to monitor SARS-CoV-2's main (M) or papain-like (PL) protease activity. These assays have proven effective in identifying potential antivirals through high-throughput screening, making fluorogenic viral protease reporters a promising platform for viral disease diagnostics and therapeutics.
SARS-CoV-2 冠状病毒通过 COVID-19 大流行在全球范围内造成了破坏，这令人清醒地提醒人们，病毒对人类健康可能产生深远影响。了解病毒的生命周期及其与宿主细胞的相互作用，为探索针对病毒相关疾病的治疗策略奠定了基础。荧光显微镜在病毒成像中发挥着至关重要的作用，提供了高时空分辨率、灵敏度和光谱多功能性。在这篇观点文章中，我们首先强调了两种最近的技术，SunTag 和 StayGold，用于病毒 RNA 翻译和病毒组装的原位成像。接下来，我们讨论了一类新的遗传编码荧光蛋白酶报告物，如 FlipGFP，它可以定制用于监测 SARS-CoV-2 的主要（M）或木瓜样（PL）蛋白酶活性。这些测定方法已被证明可通过高通量筛选有效地鉴定潜在的抗病毒药物，使荧光蛋白酶报告物成为病毒疾病诊断和治疗的有前途的平台。
