A novel codon-optimized SIV gag-pol immunogen for gene-based vaccination

Catherine M. Crosby, Eric A. Weaver, Reeti Khare, Zenaido T. Camacho, Michael A. Barry

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Simian immunodeficiency virus (SIV) is a robust pathogen used in non-human primates to model HIV vaccines. SIV encodes a number of potential vaccine targets. By far the largest and most conserved protein target in SIV is gag-pol encodes many epitopes with potential to drive multivalent T cell responses. While it is an attractive antigen, pol is only translated after a frame shift from gag so that only 1 in 10 gag proteins include this larger antigen. The codon bias of native lentiviral genes are also mismatched with the abundance of tRNAs in mammalian cells resulting in poor expression of unmodified SIV genes. To provide a better SIV gag-pol immunogen for gene-based vaccination, we codon-optimized the full gag-pol sequence from SIVmac239. To increase pol expression, we artificially moved the pol sequence in frame to gag to bypass the need for a translational frame shift for its expression. Finally, we inserted four "self-cleaving" picornavirus sequences into gag p24, protease, reverse transcriptase, and into integrase to fragment the proteins for potentially better immune presentation. We demonstrate that these immunogens are well expressed in vitro and drive similar antibody and T cell responses with or without cleavage sequences.

Original languageEnglish (US)
Pages (from-to)47-55
Number of pages9
JournalVirology Reports
Volume5
DOIs
StatePublished - Nov 25 2014
Externally publishedYes

Fingerprint

pol Genes
Simian Immunodeficiency Virus
Viruses
Codon
Vaccination
Genes
T-cells
Picornaviridae
T-Lymphocytes
gag Gene Products
Antigens
Integrases
AIDS Vaccines
RNA-Directed DNA Polymerase
Pathogens
Transfer RNA
Primates
Epitopes
Proteins
Peptide Hydrolases

Keywords

  • Adenovirus
  • Gag
  • Pol
  • SIV
  • Vaccine

ASJC Scopus subject areas

  • Biochemistry
  • Virology

Cite this

A novel codon-optimized SIV gag-pol immunogen for gene-based vaccination. / Crosby, Catherine M.; Weaver, Eric A.; Khare, Reeti; Camacho, Zenaido T.; Barry, Michael A.

In: Virology Reports, Vol. 5, 25.11.2014, p. 47-55.

Research output: Contribution to journalArticle

Crosby, Catherine M. ; Weaver, Eric A. ; Khare, Reeti ; Camacho, Zenaido T. ; Barry, Michael A. / A novel codon-optimized SIV gag-pol immunogen for gene-based vaccination. In: Virology Reports. 2014 ; Vol. 5. pp. 47-55.
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