Expression of heterologous genes in oncolytic adenoviruses using picornaviral 2A sequences that trigger ribosome skipping

The Journal of General Virology
Garth M FunstonRichard D Iggo

Abstract

Insertion of picornaviral 2A sequences into mRNAs causes ribosomes to skip formation of a peptide bond at the junction of the 2A and downstream sequences, leading to the production of two proteins from a single open reading frame. Adenoviral protein IX is a minor capsid protein that has been used to display foreign peptides on the surface of the capsid. We have used 2A sequences from the foot-and-mouth disease virus (FMDV) and porcine teschovirus 1 (PTV-1) to express protein IX (pIX) and green fluorescent protein (GFP) from pIX-2A-GFP fusion genes in an oncolytic virus derived from human adenovirus 5. GFP was efficiently expressed by constructs containing either 2A sequence. Peptide bond skipping was more efficient with the 58 aa FMDV sequence than with the 22 aa PTV-1 2A sequence, but the virus with the FMDV 2A sequence showed a reduction in plaque size, cytopathic effect, viral burst size and capsid stability. We conclude that ribosome skipping induced by 2A sequences is an effective strategy to express heterologous genes in adenoviruses; however, careful selection or optimization of the 2A sequence may be required if protein IX is used as the fusion partner.

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Citations

Mar 22, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jan Ole KemnadeDavid M Spencer
Sep 19, 2009·Nature Reviews. Genetics·D Allan Drummond, Claus O Wilke
Jul 24, 2013·BioMed Research International·Ekaterina Minskaia, Martin D Ryan
Oct 6, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Yanqing ZhangHongju Wu
Apr 19, 2011·Xenotransplantation·Nella FisicaroPeter J Cowan
Jan 1, 2010·Biotechnology & Genetic Engineering Reviews·Garry LukeMartin Ryan
Jul 28, 2020·International Journal of Molecular Sciences·Martí Farrera-SalRamon Alemany
Aug 9, 2017·Frontiers in Microbiology·Xiaoyao YangXiaoyue Chen
May 13, 2020·Nature Biotechnology·Matthew J SpindlerDavid S Johnson
Aug 12, 2021·Molecular Therapy Oncolytics·Martí Farrera-SalRamon Alemany

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