Luciferase Reporting of CrPV-Fluc mRNA

Polyadenylated CrPV-Fluc mRNA was in vitro transcribed using a 50T-tailed PCR product as a template, as described previously (Prokhorova et al. 2016 (link)). Human HEK293T cells (ATCC) were seeded in DMEM supplemented with 10% FBS onto a 96-well white opaque FB plate (Greiner). 12–16 h later, the cells were transfected with 30 ng of mRNA per well using GenJector-U (Molecta, Russia). Beetle luciferin (Promega) was added to the medium to final concentration of 0.5 mM, and the plate was immediately placed into Infinite 200 Pro microplate reader (TECAN) provided with a gas control module. The cells were incubated at 36.6°С in 5% CO₂ for 12 h, luciferase activity was continuously measured every 5 min with integration time 5 s, as describes earlier (Panova et al. 2023 (link)).