The genome position of a therapeutic transgene strongly influences the level of expression in an armed oncolytic human adenovirus vector
| Autor: | Joshua Del Papa, Ryan G. Clarkin, Kathy L. Poulin, Robin J. Parks |
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| Rok vydání: | 2020 |
| Předmět: |
Transgene
RNA Splicing Genetic Vectors Gene Expression Genome Viral Biology Genome Viral vector 03 medical and health sciences Virology Adenovirus E3 Proteins Humans Vector (molecular biology) Transgenes Gene 030304 developmental biology 0303 health sciences Adenoviruses Human 030302 biochemistry & molecular biology Adenovirus Death Protein Transmembrane protein Cell biology Oncolytic virus Oncolytic Viruses HEK293 Cells A549 Cells Viral Fusion Proteins |
| Zdroj: | Virology. 561 |
| ISSN: | 1096-0341 |
| Popis: | Efficacy of oncolytic, conditionally-replicating adenovirus (CRAd) vectors can be enhanced by “arming” the vector with therapeutic transgenes. We examined whether inclusion of an intact early region 3 (E3) and the reptilian reovirus fusogenic p14 fusion-associated small transmembrane (FAST) protein enhanced vector efficacy. The p14 FAST transgene was cloned between the fiber gene and E4 region, with an upstream splice acceptor for replication-dependent expression from the major late promoter. In A549 cells, this vector expressed p14 FAST protein at very low levels, and showed a poor ability to mediate cell-cell fusion, relative to a similar vector encoding p14 FAST within the E3 deletion. Although expression of E3 proteins from the CRAd increased plaque size, poor expression of p14 FAST protein compromised the fusogenic capacity of the vector. Thus, location of a therapeutic transgene within a CRAd can significantly impact expression of the transgene and is an important consideration in vector design. |
| Databáze: | OpenAIRE |
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