Fig. 6: Antisense targeting of highly conserved RNA secondary structures in SARS-CoV-2. | Nature Medicine

Fig. 6: Antisense targeting of highly conserved RNA secondary structures in SARS-CoV-2.

From: Programmable antivirals targeting critical conserved viral RNA secondary structures from influenza A virus and SARS-CoV-2

Fig. 6

a,b, Two SHAPE reactivity-derived prediction of RNA secondary structures within (+)-sense SARS-CoV-2 vRNA and LNAs designed against these targets: nt 28,743–28,827, LNA-12.8 (a) and nt 258–276, LNA-14.3 (b). c,d, Electrophoretic profile of SHAPE analysis performed in the presence or absence of the corresponding LNAs. e, In vitro antiviral activity of 25 nM LNAs against SARS-CoV-2-Nluc virus in Huh-7 cells (n = 4). The nucleoside analog, EIDD-1931, was included as a positive control at 5 µM and 0.5 µM. f, LNA dose–response against SARS-CoV-2-Nluc virus in ACE2-TMPRSS2-Huh-7.5 cells, in biological replicates (n = 6–7 for all treatment groups, except for NT controls where n = 12). g,h, Virus titers of supernatant collected from A549-hACE2 cells treated with LNA or EIDD-1931 followed by infection with either WT SARS-CoV-2 (g) or a patient-isolated clinical variant of SARS-CoV-2 containing multiple mutations in the spike protein region, including Asp614Gly, Glu484Lys and Asn501Tyr present in variants of concern (n = 3) (h). Statistical analysis in e, f, g and h was performed using ordinary one-way ANOVA with Dunnett’s multiple comparison tests between the controls as indicated. Error bars represent mean ± s.d.; P values are as indicated. i–k, Prevention of SARS-CoV-2 transmission in Syrian hamsters: 10-week-old female Syrian hamsters were pretreated with either 100 µg of LNA-12.8 (n = 5) or vehicle (n = 4) on day −1 and day 0 before exposure to SARS-CoV-2-infected sentinel hamsters for 2 h per day for 3 consecutive days. Four days after the initial exposure, lungs were harvested, and virus titers determined by CCID50 in triplicate: experimental timeline (i); virus titer of SARS-CoV-2 in lungs (two-sided, unpaired Student’s t-test) (j); and oropharyngeal swabs of LNA- or vehicle-treated hamsters (two-way ANOVA with multiple comparison tests against the vehicle controls) (k). The assay method detection limit is 0.7log10(CCID50). Error bars represent mean ± s.d. The panel i was created with BioRender.com.

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