Abstract
The Vero cell line is the most used continuous cell line for viral vaccine manufacturing with more than 40 years of accumulated experience in the vaccine industry. Additionally, the Vero cell line has shown a high affinity for infection by MERS-CoV, SARS-CoV, and recently SARS-CoV-2, emerging as an important discovery and screening tool to support the global research and development efforts in this COVID-19 pandemic. However, the lack of a reference genome for the Vero cell line has limited our understanding of host–virus interactions underlying such affinity of the Vero cell towards key emerging pathogens, and more importantly our ability to redesign high-yield vaccine production processes using Vero genome editing. In this paper, we present an annotated highly contiguous 2.9 Gb assembly of the Vero cell genome. In addition, several viral genome insertions, including Adeno-associated virus serotypes 3, 4, 7, and 8, have been identified, giving valuable insights into quality control considerations for cell-based vaccine production systems. Variant calling revealed that, in addition to interferon, chemokines, and caspases-related genes lost their functions. Surprisingly, the ACE2 gene, which was previously identified as the host cell entry receptor for SARS-CoV and SARS-CoV-2, also lost function in the Vero genome due to structural variations.
Highlights
Originated from a female Chlorocebus sabaeus (African GreenMonkey) kidney, the Vero cell line represents the most widely used continuous cell line for the production of viral vaccines with over 40 years of experience[1]
Vero cells have been extensively used in the current response to COVID-19 as a platform for SARS-CoV-2 isolation and replication, viral vaccine production, and identification of potential drug targets[9]
Using sequencing reads with a mean coverage per base pair of 100.2 (Fig. 1) of the African Green Monkey genome[10], we present here a principal pseudohaplotype and an alternate pseudohaplotype of the Vero genome consisting respectively of 6872 and 6876 scaffolds, with a total length of 2.9 Gb, a L50 count of 12, and NG50 length of 82 and 70 Mb (Fig. 2) with 39,449 predicted genes (29,824 genes were predicted for the African Green Monkey genome using the same default parameters), 35,004 genes and pseudo genes annotated
Summary
Monkey) kidney, the Vero cell line represents the most widely used continuous cell line for the production of viral vaccines with over 40 years of experience[1]. This includes the development and production of vaccines against dengue fever, influenza, Japanese encephalitis, polio, rabies, rotavirus, smallpox and more recently, Ebola (using a recombinant vesicular stomatitis virus)[2,3,4]. Vero cells have been extensively used in the current response to COVID-19 as a platform for SARS-CoV-2 isolation and replication, viral vaccine production, and identification of potential drug targets[9]. We propose a haplotype resolved annotated assembly of the WHO-
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