Modular nanotransporters capable of cause intracellular degradation of the N-protein of the SARS-CoV-2 virus in A549 cells with temporary expression of this protein fused with the fluorescent protein mRuby3

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Abstract

Modular nanotransporters (MNTs) have been created containing an antibody-like molecule, monobody, to the N-protein of the SARS-CoV-2 virus, as well as an amino acid sequence that attracts the E3 ligase Keap1 (E3BP). This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as the basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.

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About the authors

Y. V. Khramtsov

Institute of Gene Biology, RAS

Author for correspondence.
Email: alsobolev@yandex.ru
Russian Federation, Moscow

A. V. Ulasov

Institute of Gene Biology, RAS

Email: alsobolev@yandex.ru
Russian Federation, Moscow

T. N. Lupanova

Institute of Gene Biology, RAS

Email: alsobolev@yandex.ru
Russian Federation, Moscow

G. P. Georgiev

Institute of Gene Biology, RAS

Email: alsobolev@yandex.ru

Academician

Russian Federation, Moscow

A. S. Sobolev

Institute of Gene Biology, RAS; Lomonosov Moscow State University

Email: alsobolev@yandex.ru

Corresponding 

Russian Federation, Moscow; Moscow

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2. Fig. 1. Dependence of the relative fluorescence intensity (the fluorescence intensity before the start of thermophoresis is taken as 100%) 20 s after the start of thermophoresis on the concentration of MNT1 or cleaved MNT1 at a constant concentration of N-protein labeled with AF488 (5 nM). The standard error of relative fluorescence intensity determination is indicated (14–17 replicates).

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3. Fig. 2. Relative fluorescence of A549 cells (the fluorescence of cells to which MNT was not added was taken as 100%) when they were incubated for various times with 500 nM MNT1 or 500 nM MNT0. Mean values with corresponding standard error are shown (n = 3–9).

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