Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein
ABSTRACTThe nucleocapsid protein of SARS-CoV-2 plays significant roles in viral assembly, immune evasion, and viral stability. Due to its immunogenicity, high expression levels during COVID-19, and conservation across viral strains, it represents an attractive target for antiviral treatment. In this...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2024-04-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03410-23 |
| _version_ | 1797228862306779136 |
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| author | Yanping Huang Congcong Huang Junkai Chen Siwei Chen Bei Li Jian Li Zhixiong Jin Qiwei Zhang Pan Pan Weixing Du Long Liu Zhixin Liu |
| author_facet | Yanping Huang Congcong Huang Junkai Chen Siwei Chen Bei Li Jian Li Zhixiong Jin Qiwei Zhang Pan Pan Weixing Du Long Liu Zhixin Liu |
| author_sort | Yanping Huang |
| collection | DOAJ |
| description | ABSTRACTThe nucleocapsid protein of SARS-CoV-2 plays significant roles in viral assembly, immune evasion, and viral stability. Due to its immunogenicity, high expression levels during COVID-19, and conservation across viral strains, it represents an attractive target for antiviral treatment. In this study, we identified and characterized a single-stranded DNA aptamer, N-Apt17, which effectively disrupts the liquid-liquid phase separation (LLPS) mediated by the N protein. To enhance the aptamer’s stability, a circular bivalent form, cb-N-Apt17, was designed and evaluated. Our findings demonstrated that cb-N-Apt17 exhibited improved stability, enhanced binding affinity, and superior inhibition of N protein LLPS; thus, it has the potential inhibition ability on viral replication. These results provide valuable evidence supporting the potential of cb-N-Apt17 as a promising candidate for the development of antiviral therapies against COVID-19.IMPORTANCEVariants of SARS-CoV-2 pose a significant challenge to currently available COVID-19 vaccines and therapies due to the rapid epitope changes observed in the viral spike protein. However, the nucleocapsid (N) protein of SARS-CoV-2, a highly conserved structural protein, offers promising potential as a target for inhibiting viral replication. The N protein forms complexes with genomic RNA, interacts with other viral structural proteins during virion assembly, and plays a critical role in evading host innate immunity by impairing interferon production during viral infection. In this investigation, we discovered a single-stranded DNA aptamer, designated as N-Apt17, exhibiting remarkable affinity and specificity for the N protein. Notably, N-Apt17 disrupts the liquid-liquid phase separation (LLPS) of the N protein. To enhance the stability and molecular recognition capabilities of N-Apt17, we designed a circular bivalent DNA aptamer termed cb-N-Apt17. In both in vivo and in vitro experiments, cb-N-Apt17 exhibited increased stability, enhanced binding affinity, and superior LLPS disrupting ability. Thus, our study provides essential proof-of-principle evidence supporting the further development of cb-N-Apt17 as a therapeutic candidate for COVID-19. |
| first_indexed | 2024-04-24T15:03:26Z |
| format | Article |
| id | doaj.art-ac95edf2a27741ef83336cff617aea44 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-24T15:03:26Z |
| publishDate | 2024-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-ac95edf2a27741ef83336cff617aea442024-04-02T14:16:18ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972024-04-0112410.1128/spectrum.03410-23Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid proteinYanping Huang0Congcong Huang1Junkai Chen2Siwei Chen3Bei Li4Jian Li5Zhixiong Jin6Qiwei Zhang7Pan Pan8Weixing Du9Long Liu10Zhixin Liu11Department of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaGuangdong Provincial Key Laboratory of Virology, Institute of Medical Microbiology, Jinan University, Guangzhou, ChinaThe First Affiliated Hospital of Jinan University, Guangzhou, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaDepartment of Infectious Diseases, Renmin Hospital, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, ChinaABSTRACTThe nucleocapsid protein of SARS-CoV-2 plays significant roles in viral assembly, immune evasion, and viral stability. Due to its immunogenicity, high expression levels during COVID-19, and conservation across viral strains, it represents an attractive target for antiviral treatment. In this study, we identified and characterized a single-stranded DNA aptamer, N-Apt17, which effectively disrupts the liquid-liquid phase separation (LLPS) mediated by the N protein. To enhance the aptamer’s stability, a circular bivalent form, cb-N-Apt17, was designed and evaluated. Our findings demonstrated that cb-N-Apt17 exhibited improved stability, enhanced binding affinity, and superior inhibition of N protein LLPS; thus, it has the potential inhibition ability on viral replication. These results provide valuable evidence supporting the potential of cb-N-Apt17 as a promising candidate for the development of antiviral therapies against COVID-19.IMPORTANCEVariants of SARS-CoV-2 pose a significant challenge to currently available COVID-19 vaccines and therapies due to the rapid epitope changes observed in the viral spike protein. However, the nucleocapsid (N) protein of SARS-CoV-2, a highly conserved structural protein, offers promising potential as a target for inhibiting viral replication. The N protein forms complexes with genomic RNA, interacts with other viral structural proteins during virion assembly, and plays a critical role in evading host innate immunity by impairing interferon production during viral infection. In this investigation, we discovered a single-stranded DNA aptamer, designated as N-Apt17, exhibiting remarkable affinity and specificity for the N protein. Notably, N-Apt17 disrupts the liquid-liquid phase separation (LLPS) of the N protein. To enhance the stability and molecular recognition capabilities of N-Apt17, we designed a circular bivalent DNA aptamer termed cb-N-Apt17. In both in vivo and in vitro experiments, cb-N-Apt17 exhibited increased stability, enhanced binding affinity, and superior LLPS disrupting ability. Thus, our study provides essential proof-of-principle evidence supporting the further development of cb-N-Apt17 as a therapeutic candidate for COVID-19.https://journals.asm.org/doi/10.1128/spectrum.03410-23SARS-CoV-2nucleocapsid proteinaptamerantiviral therapy |
| spellingShingle | Yanping Huang Congcong Huang Junkai Chen Siwei Chen Bei Li Jian Li Zhixiong Jin Qiwei Zhang Pan Pan Weixing Du Long Liu Zhixin Liu Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein Microbiology Spectrum SARS-CoV-2 nucleocapsid protein aptamer antiviral therapy |
| title | Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein |
| title_full | Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein |
| title_fullStr | Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein |
| title_full_unstemmed | Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein |
| title_short | Inhibition of SARS-CoV-2 replication by a ssDNA aptamer targeting the nucleocapsid protein |
| title_sort | inhibition of sars cov 2 replication by a ssdna aptamer targeting the nucleocapsid protein |
| topic | SARS-CoV-2 nucleocapsid protein aptamer antiviral therapy |
| url | https://journals.asm.org/doi/10.1128/spectrum.03410-23 |
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