Sequencing ACE2 in Bos Taurus: A Comparison of Bovine and Human ACE 2’s interaction with SARS‐CoV‐2

Abstract

The World Health Organization’s (WHO) 2018 blueprint of priority diseases, based upon their potential to cause epidemics and public health emergencies included two respiratory illnesses, severe acute respiratory syndrome (SARS‐CoV) and middle east respiratory syndrome (MERS‐CoV), both caused by members of the Coronaviridae family (BioDefense, 2018). Now three years later, we are amid an ongoing worldwide pandemic caused by another Coronavirus, SARS‐Cov‐2. This virus has rapidly spread across the globe causing over 5.4 million deaths throughout the world. Symptoms of the viral infection include shortness of breath, fever, fatigue, muscle and body aches, headaches and most importantly acute lung injury (ALI) caused by dysregulation of pulmonary RAS (Renin‐Angiotensin System). RAS, the main network controlling blood pressure is regulated by ACE (Angiotensin Converting Enzyme) and its homolog ACE2. It is now known that the SARS‐CoV‐2, uses ACE2 as the cell receptor to enter and infiltrate the host cell. While ACE promotes atrophy, fibrosis, inflammation, oxidants, and vasoconstriction, ACE2 through its metabolism of angiotensin I, restricts excessive atrophy, fibrosis and causes vasodilation. Furthermore, ACE2 with its anti‐inflammatory and antioxidant role protects lung tissues and therefore a downregulation of ACE2 by SARS‐Cov‐2 infection can result in development of acute respiratory distress syndrome (ARDS) (Imai, Kuba, & Penninger, 2008). The Spike (S) protein of SARS‐Cov‐2 binds to ACE2, and recent studies have identified several residues in Exon 1 of ACE‐2, that mediate this binding (Rehman & Tabish, 2020). Our study aims to sequence ACE2 Exon 1 from Bos taurus and then compare it to human ACE2 Exon 1 using a variety of bioinformatics tools to understand and possibly identify polymorphisms that could determine species susceptibility to Covid‐19 infection. This investigation began with genomic DNA extraction from calf thymus. The genomic DNA was then analyzed using spectrophotometry (Biophotometer and Nanodrop) and gel electrophoresis. Thereafter, ACE2 target primers were designed using Primer BLAST and Primer Quest. We are now getting ready to run PCR to amplify our ACE2 Exon 1 target from Bos taurus, and after purifying the PCR products, we will sequence the ACE2 Exon 1 using our SeqStudio genetic analyzer. The sequence will then be analyzed and compared to its human counterpart using a variety of bioinformatics tools to gain a better understanding of polymorphisms that might influence SARS‐Cov‐2’s initial interaction with its host as well as pathogenesis of COVID‐19 in general.ReferencesBioDefense, G. (2018, February 12). https://globalbiodefense.com/2018/02/12/who‐updates‐blueprint‐list‐of‐priority‐diseases/. Retrieved May 14, 2020Imai, Y., Kuba, K., & Penninger, J. M. (2008). The discovery of angiotensin‐converting enzyme 2 and its role in acute lung injury in mice. Exp Physiol 93.5, pp 543–548.Rehman, S. u., & Tabish, M. (2020). Alternative splicing of ACE2 possibly generates variants that may limit the entry of SARS‐CoV‐2: a potential therapeutic approach using SSOs. Clinical Science, 134, 1143–1150.