Monmouth University East Brunswick , New Jersey, United States
Flobater Gawargi (Monmouth University)| Laura Sine (Monmouth University)| Martin Hicks (Monmouth University)
Emerging viral diseases have increased in recent decades. In December 2019, an epidemic with low respiratory infections emerged in Wuhan, China. The disease, Covid-19 was found to be caused by a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of January 07, 2021, WHO has confirmed 88,005,213 global cases and 1,897,568 deaths worldwide, 365,174 in the USA. Fortunately, a vaccine has recently been approved, yet there are no therapeutics for infected individuals. From advances in biotechnology, the genome and structure of SARS-CoV-2 is known. Three proteins are anchored in the viral envelope, Spike (S), Envelope (E), and Membrane (M), which is linked to the Nucleocapsid (N) protein connecting to the viral RNA genome. Our lab is developing an innovative therapy that delivers multiple therapeutic microRNAs to simultaneously block the expression of these distinct viral proteins. In the current work, we propose an anti-Covid microRNA therapy designed to degrade each of the mRNA transcripts of these critical genes, stopping viral assembly, and reducing the severity of infection. The design of the anti-Covid microRNAs 1) mimics human microRNA cluster 17-92a structural stability, 2) forms guide-RNA substrates for the RNA induced silencing complex, and 3) are complementary to specific regions of the SARS-CoV-2 RNA genome without off-targets effects in the human genome. Twenty-one microRNA sequences were designed to target the S gene, six for N, two for M, and one for E. These were cloned into our microRNA-17-92 therapy vector which expresses six distinct anti-Covid RNA therapeutics simultaneously. We have stably transfected the S and N gene into our tissue culture model to measure the efficacy of the anti-Covid microRNA therapy to down-regulate the S and N protein expression. We will be presenting on the efficacy of the anti-Covid microRNA to knockdown the expression of the viral proteins, S and N. In addition, we are examining the secondary structure of our RNA therapy using SHAPE-MAP to optimize RNA therapeutic stability in comparison to the stable structure of the original Mir-17-92a.