SARS-Coronavirus-2 Gene Mutagenesis
Abstract
Coronavirus disease 19 (COVID-19) is a respiratory disease that is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Beginning in late 2019, infection and transmission of SARS-CoV-2 has led to a global pandemic that continues to be a major public health problem resulting in more than 6 million deaths worldwide. SARS-CoV-2 binding is mediated by the Spike (S) attachment protein. The S protein is composed of two subunits, S1 and S2. The S1 subunit houses the receptor binding domain (RBD) that is responsible for recognizing and binding to the host viral receptor angiotensin-converting enzyme 2 (ACE2). The S2 subunit is responsible for fusion between viral and host cell membranes. Because of its importance in virus binding and infection, the S protein has been the target for vaccine development. For example, neutralizing antibodies, induced by the SARS-CoV-2 vaccine generally bind to the S protein blocking ACE2 binding and aborting virus infection. However, the SARS-CoV-2 virus continues to evolve, accumulating mutations in the S protein that result in immune evasion and/or increased transmission. For example, S protein L452R, T478K, D614G, P681R, and D950N mutations are associated with increased infectivity, evasion of host immune functions, and enhanced disease. To tease out the functions of each and multiple combinations of these mutations we mutated the alpha variant S protein incorporating random combinations of L452R, T478K, D614G, P681R, and D950N mutations. This was done by introducing primers of differing concentrations to generate a library of transformants harboring unique combinations of S protein mutations. Sequencing of these transformants to confirm mutations is currently pending. The future goal of this project is to use safe-to-use virus-like particles psuedotyped with the mutated SARS-CoV-2 S proteins which will be used in entry and immune cell deregulation assays.
College
College of Science & Engineering
Department
Biology
Campus
Winona
First Advisor/Mentor
Osvaldo Martinez
Start Date
4-19-2023 9:00 AM
End Date
4-19-2023 10:00 AM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Poster Number
28
SARS-Coronavirus-2 Gene Mutagenesis
Coronavirus disease 19 (COVID-19) is a respiratory disease that is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Beginning in late 2019, infection and transmission of SARS-CoV-2 has led to a global pandemic that continues to be a major public health problem resulting in more than 6 million deaths worldwide. SARS-CoV-2 binding is mediated by the Spike (S) attachment protein. The S protein is composed of two subunits, S1 and S2. The S1 subunit houses the receptor binding domain (RBD) that is responsible for recognizing and binding to the host viral receptor angiotensin-converting enzyme 2 (ACE2). The S2 subunit is responsible for fusion between viral and host cell membranes. Because of its importance in virus binding and infection, the S protein has been the target for vaccine development. For example, neutralizing antibodies, induced by the SARS-CoV-2 vaccine generally bind to the S protein blocking ACE2 binding and aborting virus infection. However, the SARS-CoV-2 virus continues to evolve, accumulating mutations in the S protein that result in immune evasion and/or increased transmission. For example, S protein L452R, T478K, D614G, P681R, and D950N mutations are associated with increased infectivity, evasion of host immune functions, and enhanced disease. To tease out the functions of each and multiple combinations of these mutations we mutated the alpha variant S protein incorporating random combinations of L452R, T478K, D614G, P681R, and D950N mutations. This was done by introducing primers of differing concentrations to generate a library of transformants harboring unique combinations of S protein mutations. Sequencing of these transformants to confirm mutations is currently pending. The future goal of this project is to use safe-to-use virus-like particles psuedotyped with the mutated SARS-CoV-2 S proteins which will be used in entry and immune cell deregulation assays.
