• Design of multi-antigen vaccine based on mRNA platform against cytomegalovirus and evaluation of expression of vaccine antigens at protein level in Vitro
  • Mohammad Hossein. Nicknam ,1 Massoud Soleimani ,2,* Somayeh Mami,3 Sajjad shekarchain ,4
    1. Department of Immunology, molecular immunology research center,Tehran University of Medical Sciences
    2. Department of Hematology,School of Medical Sciences, Tarbiat Modares University
    3. Department of Immunology, Tehran University of Medical Sciences
    4. Department of Immunology, Shahed University, Tehran, Iran.


  • Introduction: Cytomegalovirus is a widespread infection that is prevalent globally. In healthy individuals, this infection typically remains asymptomatic due to its slow replication, immune system inhibition, and suppression, but persists in the body indefinitely following initial infection. Mononucleosis is the most common complication resulting from this virus in healthy individuals. However, cytomegalovirus is considered a hazardous pathogen for those with weakened or suppressed immune systems, including organ transplant recipients, AIDS patients, premature infants, and individuals with various malignancies. At present, there is no approved vaccine available against CMV. Given the recent surge in pharmaceutical sanctions and the associated challenges and costs associated with accessing vaccines produced abroad, it would be difficult and expensive for the people of Iran to obtain such a vaccine. In light of these circumstances and the paramount importance of this subject matter, our objective in this study is to develop an mRNA vaccine against cytomegalovirus.
  • Methods: Initially, the desired antigens were meticulously selected by thoroughly scrutinizing scientific sources and utilizing immunoinformatic methods. Subsequently, an appropriate genetic construct was designed to express the chosen antigens, followed by the design of a suitable clonal vector for the genetic construct. The vector was then procured from Gene Universal for synthesis. Upon receipt of the designed vector, the appropriate cell was prepared and the vector was transformed into it. Bacteria containing the desired vector were then identified and cultured. The cloned plasmid was extracted and linearized using the appropriate restriction enzyme. An IVT reaction was performed using linearized DNAs. Thereafter, if the desired mRNA was properly synthesized, it was extracted and labeled. The protein expression of mRNAs in specific cell lines was measured using the western blot method.
  • Results: Upon receipt of the intended plasmids and their subsequent cloning in bacterial cells, it was observed that the insert sequence had been correctly integrated. Subsequently, these plasmids was subjected to purification and concentration measurement via nanodrop, rendering them suitable for employment in the in vitro transcription (IVT) reaction. After purification and encapsulation of mRNA and their treatment on cell lines and performing western blot and Flow cytometry , it was observed that the proteins had an acceptable expression.
  • Conclusion: According to the obtained results, it is concluded that the synthesized mRNAs have the ability to be expressed into protein and create antigens related to the cytomegalovirus virus, which probably has the potential to stimulate the immune system.
  • Keywords: Cytomegalovirus, mRNA vaccine