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Reverse genetics allows for the molecular study of pathogenic determinants to enable rational vaccine design. The recombinant IBV (rIBV) Beau-R, a molecular clone of the apathogenic Beaudette strain, has previously been investigated as a vaccine platform. To determine tissues in which Beau-R could effectively deliver antigenic genes, an in vivo study in chickens, the natural host, was used to compare the pattern of viral dissemination of Beau-R to the pathogenic strain M41-CK. Replication of Beau-R was found to be restricted to soft tissue within the beak, whereas M41-CK was detected in beak tissue, trachea and eyelid up to seven days post infection. In vitro assays further identified that, unlike M41-CK, Beau-R could not replicate at 41 \u00b0C, the core body temperature of a chicken, but is able to replicate a 37 \u00b0C, a temperature relatable to the very upper respiratory tract. Using a panel of rIBVs with defined mutations in the structural and accessory genes, viral replication at permissive and non-permissive temperatures was investigated, identifying that the Beau-R replicase gene was a determinant of temperature sensitivity and that sub-genomic mRNA synthesis had been affected. The identification of temperature sensitive allelic lesions within the Beau-R replicase gene opens up the possibility of using this method of attenuation in other IBV strains for future vaccine development as well as a method to investigate the functions of the IBV replicase proteins.<\/jats:p>","DOI":"10.3390\/v12070754","type":"journal-article","created":{"date-parts":[[2020,7,14]],"date-time":"2020-07-14T11:03:23Z","timestamp":1594724603000},"page":"754","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus"],"prefix":"10.3390","volume":"12","author":[{"given":"Sarah","family":"Keep","sequence":"first","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Phoebe","family":"Stevenson-Leggett","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Steyn","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael S.","family":"Oade","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isobel","family":"Webb","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamie","family":"Stuart","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2241-1743","authenticated-orcid":false,"given":"Lonneke","family":"Vervelde","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, The Roslin Institute and Royal (Dick), School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Britton","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2901-5578","authenticated-orcid":false,"given":"Helena J.","family":"Maier","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4012-1283","authenticated-orcid":false,"given":"Erica","family":"Bickerton","sequence":"additional","affiliation":[{"name":"The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.1126\/science.1085952","article-title":"Characterization of a novel coronavirus associated with severe acute respiratory syndrome","volume":"300","author":"Rota","year":"2003","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1128\/MMBR.69.4.635-664.2005","article-title":"Coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus","volume":"69","author":"Weiss","year":"2005","journal-title":"Microbiol. 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