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Research Article | Open Access

RNAi Degrades the SARS-CoV-2 Spike Protein RNA for Developing Drugs to Treat COVID-19

Weiwei Zhang1,2,3, Linjia Huang2, Jumei Huang2, Xin Jiang3,4, Xiaohong Ren6, Xiaojie Shi1, Ling Ye2,3, Shuhui Bian4, Jianhe Sun2, Yufeng Gao2, Zehua Hu2, Lintin Guo2, Suyan Chen4, Jiahao Xu2, Jie Wu1,3, Jiwen Zhang3,6( ), Daxiang Cui5( ), Fangping Dai1,2,3,5( )
Central Laboratory, Nantong Haimen People's Hospital, Nantong 226199, China
Genome-decoding Biomedical Technology Co., Ltd., Nantong 226126, China
Yangtze Delta Drug Advanced Research Institute, Nantong 226126, China
Precision-genes Bio-Technology Co., Ltd/Medical Laboratory, Nantong 226126, China
National Engineering Research Center for Nanotechnology, Shanghai Jiao Tong University, Shanghai 200241, China
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
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Abstract

COVID-19 is caused by severe acute respiratory SARS-CoV-2. Regardless of the availability of treatment strategies for COVID-19, effective therapy will remain essential. A promising approach to tackle the SARS-CoV-2 could be small interfering (si) RNAs. Here we designed the small hairpin RNA (named as shRNA688) for targeting the prepared 813 bp Est of the S protein genes (Delta). The conserved and mutated regions of the S protein genes from the genomes of the SARS-CoV-2 variants in the public database were analyzed. A 813 bp fragment encoding the most part of the RBD and partial downstream RBD of the S protein was cloned into the upstream red florescent protein gene (RFP) as a fusing gene in the pCMV-S-Protein RBD-Est-RFP plasmid for expressing a potential target for RNAi. The double stranded of the DNA encoding for shRNA688 was constructed in the downstream human H1 promoter of the plasmid in which CMV promoter drives enhanced green fluorescent protein (EGFP) marker gene expression. These two kinds of the constructed plasmids were co-transfected into HEK293T via Lipofectamine 2000. The degradation of the transcripts of the SARS-CoV-2 S protein fusing gene expressed in the transfected HEK293T treated by RNAi was analyzed by RT-qPCR with a specific probe of the targeted SARS-CoV-2 S protein gene transcripts. Our results showed that shRNA688 targeting the conserved region of the S protein genes could effectively reduce the transcripts of the S protein genes. This study provides a cell model and technical support for the research and development of the broad-spectrum small nucleic acid RNAi drugs against SARS-CoV-2 or the RNAi drugs for the other hazard viruses which cause human diseases.

Keywords

COVID-19RNAiNanoparticlesshRNAPlasmids

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Nano Biomedicine and Engineering
Volume 14 Issue 2,
June 2022
Pages 173-185
DOI: 10.5101/nbe.v14i2.p173-185
Cite this article:
Zhang W, Huang L, Huang J, et al. RNAi Degrades the SARS-CoV-2 Spike Protein RNA for Developing Drugs to Treat COVID-19. Nano Biomedicine and Engineering, 2022, 14(2): 173-185. https://doi.org/10.5101/nbe.v14i2.p173-185

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Received: 21 October 2022
Accepted: 07 November 2022
Published: 07 November 2022
© Weiwei Zhang, Linjia Huang, Jumei Huang, Xin Jiang, Xiaohong Ren, Xiaojie Shi, Ling Ye, Shuhui Bian, Jianhe Sun, Yufeng Gao, Zehua Hu, Lintin Guo, Suyan Chen, Jiahao Xu, Jie Wu, Jiwen Zhang, Daxiang Cui, and Fangping Daii.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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