High-level expression of rabies virus glycoprotein with the RNA-based Semliki Forest Virus expression vector.
Rabies is to this date one of the most important death causing zoonotic viral diseases, with 98% of deaths reported in developing countries, where access to modern vaccines and tools for efficient diagnostic remain unaffordable. In this paper, we describe a newly engineered RNA-based rabies virus glycoprotein (RVGP) expression vector based on the Semliki Forest Virus (SFV) system. A recombinant SFV carrying an RNA coding for RVGP (SFV-RVGP) was constructed and the RVGP expression was evaluated in animal cell cultures. The mRNA coding for RVGP and the RVGP itself were assessed by qPCR, Western-blotting, confocal microscopy, flow cytometry and ELISA. Moreover, SFV-RVGP was proven to be highly efficient in expressing the functionally trimeric RVGP. SFV-RVGP is easy to produce and efficient in different cell lines, making it an interesting candidate for efficient and functional viral glycoprotein expression.
Benmaamar
Rim
R
Dpt Récepteurs et Protéines Membranaires, CNRS-UMR7175, Illkirch, France.
Astray
Renato Mancini
RM
Wagner
Renaud
R
Pereira
Carlos Augusto
CA
D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D012367:RNA, Viral / Q000096:biosynthesis*; D011820:Rabies virus / Q000235:genetics*; D012672:Semliki forest virus / Q000235:genetics*; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*
D006023:Glycoproteins; D012367:RNA, Viral; D014764:Viral Proteins