{"doi":"10.1074/jbc.ra120.016148","title":"SARS-CoV-2 viral budding and entry can be modeled using BSL-2 level virus-like particles","abstract":"Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first discovered in December 2019 in Wuhan, China, and expeditiously spread across the globe causing a global pandemic. Research on SARS-CoV-2, as well as the closely related SARS-CoV-1 and MERS coronaviruses, is restricted to BSL-3 facilities. Such BSL-3 classification makes SARS-CoV-2 research inaccessible to the majority of functioning research laboratories in the United States; this becomes problematic when the collective scientific effort needs to be focused on such in the face of a pandemic. However, a minimal system capable of recapitulating different steps of the viral life cycle without using the virus’ genetic material could increase accessibility. In this work, we assessed the four structural proteins from SARS-CoV-2 for their ability to form virus-like particles (VLPs) from human cells to form a competent system for BSL-2 studies of SARS-CoV-2. Herein, we provide methods and resources of producing, purifying, fluorescently and APEX2-labeling of SARS-CoV-2 VLPs for the evaluation of mechanisms of viral budding and entry as well as assessment of drug inhibitors under BSL-2 conditions. These systems should be useful to those looking to circumvent BSL-3 work with SARS-CoV-2 yet study the mechanisms by which SARS-CoV-2 enters and exits human cells. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first discovered in December 2019 in Wuhan, China, and expeditiously spread across the globe causing a global pandemic. Research on SARS-CoV-2, as well as the closely related SARS-CoV-1 and MERS coronaviruses, is restricted to BSL-3 facilities. Such BSL-3 classification makes SARS-CoV-2 research inaccessible to the majority of functioning research laboratories in the United States; this becomes problematic when the collective scientific effort needs to be focused on such in the face of a pandemic. However, a minimal system capable of recapitulating different steps of the viral life cycle without using the virus’ genetic material could increase accessibility. In this work, we assessed the four structural proteins from SARS-CoV-2 for their ability to form virus-like particles (VLPs) from human cells to form a competent system for BSL-2 studies of SARS-CoV-2. Herein, we provide methods and resources of producing, purifying, fluorescently and APEX2-labeling of SARS-CoV-2 VLPs for the evaluation of mechanisms of viral budding and entry as well as assessment of drug inhibitors under BSL-2 conditions. These systems should be useful to those looking to circumvent BSL-3 work with SARS-CoV-2 yet study the mechanisms by which SARS-CoV-2 enters and exits human cells. Severe acute respiratory syndrome SARS-coronavirus 2 (SARS-CoV-2) emerged in December 2019 in Wuhan, China, and has since spread around the globe. As of late October 2020, the virus has been detected in 189 different countries and territories with more than 43 million confirmed cases and more than one million attributed fatalities (https://covid19.who.int). The virulence of coronaviruses has previously been observed in severe acute respiratory syndrome coronavirus (SARS-CoV-1) and Middle East Respiratory Syndrome coronavirus (MERS-CoV) outbreaks in the previous 2 decades; however, there still remains no FDA-approved treatment for any coronavirus. In order to develop therapeutics, the ability to study viruses must be accessible. Under current circumstances, the authentic live SARS-CoV-2 virus is restricted to BSL-3 containment facilities; while many of these facilities exist and have refocused their collective efforts on SARS-CoV-2 research, the greater scientific community could be vastly helpful in combating this pandemic if such research was BSL-2 compatible. For instance, BSL-2 models of other difficult-to-work-with BSL-3 and -4 pathogens such as SARS-CoV-1 (1Siu Y.L. Teoh K.T. Lo J. Chan C.M. Kien F. Escriou N. Tsao S.W. Nicholls J.M. Altmeyer R. Peiris J.S. Bruzzone R. Nal B. The M, E, and N ","journal":"Journal of Biological Chemistry","year":2020,"id":53326,"datarank":0.7181237614173069,"base_score":4.787491742782046,"endowment":4.787491742782046,"self_citation_contribution":0.7181237614173069,"citation_network_contribution":0.0,"self_endowment_contribution":0.7181237614173069,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":119,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.959,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":271700,"name":"Emily A. David","orcid":"0000-0002-1106-0725","position":1,"is_corresponding":false},{"id":271701,"name":"Dhabaleswar Patra","orcid":"0000-0001-6520-6972","position":2,"is_corresponding":false},{"id":271702,"name":"Ranjan Sengupta","orcid":"0000-0003-3160-5312","position":3,"is_corresponding":false},{"id":271703,"name":"Souad Amiar","orcid":"0000-0002-6117-0746","position":4,"is_corresponding":false},{"id":271704,"name":"Yuan Su","orcid":"0000-0003-4406-3946","position":5,"is_corresponding":false},{"id":271705,"name":"Robert V. Stahelin","orcid":"0000-0001-5443-7863","position":6,"is_corresponding":false},{"id":271699,"name":"Caroline B. Plescia","orcid":"0000-0003-0520-4937","position":0,"is_corresponding":true}],"reference_count":28,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T20:44:25.349797Z","pmid":"33214224","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}