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SCIENCE CHINA Life Sciences, Volume 63 , Issue 9 : 1413-1416(2020) https://doi.org/10.1007/s11427-020-1692-1

SARS-CoV-2-encoded nucleocapsid protein acts as a viral suppressor of RNA interference in cells

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  • ReceivedFeb 23, 2020
  • AcceptedMar 4, 2020
  • PublishedApr 10, 2020

Abstract

There is no abstract available for this article.


Funded by

the Strategic Priority Research Program of CAS(XDB29010300,to,X.Z.)

the National Natural Science Foundation of China(31800140,to,J.M.,81873964,to,Y.Q.,31670161,to,X.Z.)

the National Science and Technology Major Project(2018ZX10101004,to,X.Z.)

and the Yunde Hou Academician Fund from National Institute for Viral Disease Control and Prevention(2019HYDQNJJ10,to,J.M.)


Acknowledgment

We thank Prof. Peng Zhou (Wuhan, China) for kindly providing materials and thank Yan Wu and Weijuan Shang for experimental assistance. This work was supported by the Strategic Priority Research Program of CAS (XDB29010300 to X.Z.), the National Natural Science Foundation of China (31800140 to J.M., 81873964 to Y.Q., and 31670161 to X.Z.), the National Science and Technology Major Project (2018ZX10101004 to X.Z.), and the Yunde Hou Academician Fund from National Institute for Viral Disease Control and Prevention (2019HYDQNJJ10 to J.M.).


Interest statement

The author(s) declare that they have no conflict of interest.


Supplement

SUPPORTING INFORMATION

Supplementary Materials and Methods

Figure S1 Schematic diagram of N proteins of SARS-CoV-2 (GISAID, accession number: EPI_ISL_402124) and different SARS-CoV strains (NCBI, accession number: NC_004718, AY502924 and AY536760).

Table S1 The primers and oligonucleotides used in this study

The supporting information is available online at http://life.scichina.com and https://link.springer.com. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.


References

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  • Figure 1

    SARS-CoV-2 N suppressed RNAi in cultured human cells. HEK293T cells were co-transfected with plasmids encoding EGFP (0.1 µg) and EGFP-specific shRNA (0.3 µg), together with either empty vector or the plasmid encoding SARS-CoV-2 N or EBOV VP35 (2 µg each). A, At 48 hpt, cells were observed via fluorescent microscopy. Scale bar, 400 µm. B, Cell lysates were harvested and analyzed by Western blotting with anti-EGFP, anti-FLAG and anti-Tubulin antibodies. C, Total RNAs were extracted and EGFP mRNA levels were examined by Northern blotting with a DIG-labeled RNA probe targeting EGFP ORF 1–400 nt. 18S and 28S rRNAs were used as loading controls. D, HEK293T cells were transfected with the plasmid encoding SARS-CoV-2 N or empty vector, together with EGFP-specific dsRNA. At 24 hpt, the cell lysates were subjected to RNA-IP with anti-FLAG or anti-IgG antibodies. Input and precipitated RNAs and proteins were detected by Northern blotting and Western blotting, respectively. HEK293T cells were co-transfected with the plasmid encoding EGFP (0.1 µg) and EGFP-specific siRNA (0.3 µg), together with either empty vector or the plasmid encoding SARS-CoV-2 N or EBOV VP35 (2 µg each). E, At 48 hpt, cells were observed via fluorescent microscopy. Scale bar, 400 µm. F, Cell lysates were harvested and analyzed by Western blotting with anti-EGFP, anti-FLAG and anti-Tubulin antibodies. G, Total RNAs were extracted and EGFP mRNA levels were examined by Northern blotting. 18S and 28S rRNAs were used as loading controls.

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