{"doi":"10.3389/fcimb.2023.1268227","title":"Cell type dependent stability and virulence of a recombinant SARS-CoV-2, and engineering of a propagation deficient RNA replicon to analyze virus RNA synthesis","abstract":"Engineering of reverse genetics systems for newly emerged viruses allows viral genome manipulation, being an essential tool for the study of virus life cycle, virus-host interactions and pathogenesis, as well as for the development of effective antiviral strategies. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emergent human coronavirus that has caused the coronavirus disease (COVID-19) pandemic. The engineering of a full-length infectious cDNA clone and a fluorescent replicon of SARS-CoV-2 Wuhan-Hu-1, using a bacterial artificial chromosome, is reported. Viral growth and genetic stability in eleven cell lines were analyzed, showing that both VeroE6 cells overexpressing transmembrane serin protease 2 (TMPRSS2) and human lung derived cells resulted in the optimization of a cell system to preserve SARS-CoV-2 genetic stability. The recombinant SARS-CoV-2 virus and a point mutant expressing the D614G spike protein variant were virulent in a mouse model. The RNA replicon was propagation-defective, allowing its use in BSL-2 conditions to analyze viral RNA synthesis. The SARS-CoV-2 reverse genetics systems developed constitute a useful tool for studying the molecular biology of the virus, the development of genetically defined vaccines and to establish systems for antiviral compounds screening.","journal":"Frontiers in Cellular and Infection Microbiology","year":2023,"id":359861,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":10,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9668,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1110833,"name":"María Adelina Schlie Guzmán","orcid":"0000-0002-9461-2547","position":1,"is_corresponding":false},{"id":935878,"name":"Diego Muñoz‐Santos","orcid":"0000-0003-0591-0874","position":2,"is_corresponding":false},{"id":713150,"name":"José M. Honrubia","orcid":"0000-0003-1851-1143","position":3,"is_corresponding":false},{"id":935881,"name":"Jorge Ripoll-Gómez","orcid":"0000-0003-2904-7876","position":4,"is_corresponding":false},{"id":1110834,"name":"Rafaël Delgado","orcid":"0000-0002-6912-4736","position":5,"is_corresponding":false},{"id":306889,"name":"Isabel Sola","orcid":"0000-0002-5704-1917","position":6,"is_corresponding":false},{"id":361115,"name":"Luis Enjuanes","orcid":"0000-0002-0854-0226","position":7,"is_corresponding":false},{"id":411048,"name":"Sonia Zúñiga","orcid":"0000-0003-2549-6826","position":8,"is_corresponding":false},{"id":411046,"name":"Li Wang","orcid":"0000-0001-9639-1058","position":0,"is_corresponding":true}],"reference_count":113,"raw_metadata":null,"created_at":"2026-07-19T01:13:57.361401Z","pmid":"37942479","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":[]}