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Enhancement of Transgene Production in a Vaccinia Virus‐based Expression System
Author(s) -
Richard Brent,
Flores Julia,
Jacobs Bertram,
Chaput John,
Wellensiek Brian
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.600.13
Subject(s) - vaccinia , virology , transgene , biology , immune system , modified vaccinia ankara , vaccination , attenuated vaccine , population , virus , gene , immunology , recombinant dna , genetics , medicine , virulence , environmental health
Poxvirus‐based vaccinations, i.e., vaccinia viruses (VACV), have been used more extensively for human immunization than any other vaccine in human history. VACV‐based vaccines are of particular interest due to the relative ease with which the VACV genome can be modified to express genes from a variety of sources. In recent years, there have been great advances in VACV‐based vaccines, including the generation of replication‐competent (non‐attenuated) and replication‐incompetent (attenuated) strains of VACV. While non‐attenuated VACV are highly immunogenic due to their ability to replicate in human cells, vaccines generated using these strains have higher than normal complication rates and are restricted in the population that can receive them. Attenuated strains lack the ability to replicate in human cells, resulting in a dramatic decrease in side effects, however this increase in safety comes at the cost of a reduced immune response. To address this issue, our current research is focused on increasing viral protein production in an attenuated VACV. In previous studies, we have discovered a human translation enhancing element (hTEE‐658) capable of serving as a VACV promoter. Characterization of this sequence has demonstrated that when this element is used in a VACV‐based expression system, it leads to increased protein expression levels over those driven by standard VACV promoters. Further optimization of expression levels driven by hTEE‐658, through varying the length of the transgene leader sequence, has resulted in the generation of a sequence element capable of driving high levels of protein production in an attenuated VACV system. These results provide a promising option for the production of safe, highly immunogenic VACVs that can be used to develop vaccines against a wide range of infectious diseases. Support or Funding Information Funding for this research was provided by Midwestern University intramural funds awarded to BPW and an NIH grant awarded to JCC