@article{51ad47d2c1474b16928c14cea2883319,
title = "In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome",
abstract = "Cytomegaloviruses (CMVs) are highly adapted to their host species resulting in strict species specificity. Hence, in vivo examination of all aspects of CMV biology employs animal models using host-specific CMVs. Infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a representative model for infection of humans with HCMV due to the close evolutionary relationships of both host and virus. However, the only available RhCMV clone that permits genetic modifications is based on the 68-1 strain which has been passaged in fibroblasts for decades resulting in multiple genomic changes due to tissue culture adaptations. As a result, 68-1 displays reduced viremia in RhCMV-na{\"i}ve animals and limited shedding compared to non-clonal, low passage isolates. To overcome this limitation, we used sequence information from primary RhCMV isolates to construct a full-length (FL) RhCMV by repairing all mutations affecting open reading frames (ORFs) in the 68-1 bacterial artificial chromosome (BAC). Inoculation of adult, immunocompetent, RhCMV-na{\"i}ve RM with the reconstituted virus resulted in significant viremia in the blood similar to primary isolates of RhCMV and furthermore led to high viral genome copy numbers in many tissues at day 14 post infection. In contrast, viral dissemination was greatly reduced upon deletion of genes also lacking in 68-1. Transcriptome analysis of infected tissues further revealed that chemokine-like genes deleted in 68-1 are among the most highly expressed viral transcripts both in vitro and in vivo consistent with an important immunomodulatory function of the respective proteins. We conclude that FL-RhCMV displays in vitro and in vivo characteristics of a wildtype virus while being amenable to genetic modifications through BAC recombineering techniques.",
author = "Husam Taher and Eisa Mahyari and Craig Kreklywich and Uebelhoer, {Luke S.} and McArdle, {Matthew R.} and Mostr{\"o}m, {Matilda J.} and Amruta Bhusari and Michael Nekorchuk and E. Xiaofei and Travis Whitmer and Scheef, {Elizabeth A.} and Sprehe, {Lesli M.} and Roberts, {Dawn L.} and Hughes, {Colette M.} and Jackson, {Kerianne A.} and Selseth, {Andrea N.} and Ventura, {Abigail B.} and Cleveland-Rubeor, {Hillary C.} and Yujuan Yue and Schmidt, {Kimberli A.} and Jason Shao and Edlefsen, {Paul T.} and Jeremy Smedley and Kowalik, {Timothy F.} and Stanton, {Richard J.} and Axthelm, {Michael K.} and Estes, {Jacob D.} and Hansen, {Scott G.} and Amitinder Kaur and Barry, {Peter A.} and Bimber, {Benjamin N.} and Picker, {Louis J.} and Streblow, {Daniel N.} and Klaus Fr{\"u}h and Daniel Malouli",
note = "Funding Information: National Institute of Allergy and Infectious Diseases (NIAID) https://www.niaid.nih.gov/: P01 AI129859U42 PAB, AK, KF R01 AI095113 to LJP P01 AI094417 to LJP R37 AI054292 to LJP R01 AI059457 to KF OD023038 to MKA the National Institutes of Health Office of the Director https:// www.nih.gov/about-nih/what-we-do/nih-almanac/ office-director-nih: U42OD010426 to OHSU P51OD011092 to OHSU P51OD011104 to Tulane University the Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD) https://www.nichd.nih.gov/: 4DP2HD075699 to AK the Bill & Melinda Gates Foundation https://www.gatesfoundation.org/: OPP1033121 to LJP OPP1108533 to LJP OPP1152430 to LJP The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: Copyright: {\textcopyright} 2020 Taher et al. 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.",
year = "2020",
month = nov,
day = "24",
doi = "10.1371/journal.ppat.1008666",
language = "English (US)",
volume = "16",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "11",
}