Extended transgene expression from a nonintegrating adenoviral vector containing retroviral elements

Changyu Zheng, Joseph M Vitolo, Weitian Zhang, Fumi Mineshiba, John A. Chiorini, Bruce J. Baum

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We studied the effects of specific retroviral elements in a first-generation serotype 5 adenoviral (Ad5) vector, AdLTR2 EF1α-hEPO. This vector contains 858 base pair (bp) [251-bp envelope sequence plus 607-bp long-terminal repeat (LTR)] from Moloney murine leukemia virus (MoMLV), upstream of the human elongation factor-1α (EF1α) promoter and human erythropoietin (hEPO) cDNA, with the LTR sequence downstream of the polyadenylation signal. We compared expression of AdLTR2EF1α-hEPO with corresponding expressions of two conventional Ad5 vectors, AdEF1α-hEPO and AdCMV-hEPO, in vivo in submandibular glands in rats. Both the conventional vectors yielded low serum hEPO levels by day 7, and little change in hematocrits. In contrast, after receiving AdLTR2EF1α-hEPO, the rats showed elevated hEPO levels and hematocrits for 1-3 months. In vitro studies showed that the integration efficiencies of all the vectors were similar (∼10-3). Approximately 0.1% of the vector genomes were present 1 year after delivery in the case of each of the three vectors, primarily as intact linear double-strand DNA. The unique results seen with AdLTR2EF1α-hEPO are partly because of LTR enhancer activity. However, other cis-acting activity, which is not immunomodulatory but nevertheless influences promoter methylation, appears to be involved. A vector such as AdLTR2 EF1α-hEPO may prove useful in clinical applications in which extended, but not "permanent," transgene expression is desirable.

Original languageEnglish (US)
Pages (from-to)1089-1097
Number of pages9
JournalMolecular Therapy
Volume16
Issue number6
DOIs
StatePublished - Jan 1 2008
Externally publishedYes

Fingerprint

Transgenes
Erythropoietin
Terminal Repeat Sequences
Peptide Elongation Factor 1
Base Pairing
Hematocrit
Moloney murine leukemia virus
Polyadenylation
Submandibular Gland
Methylation
Complementary DNA
Genome
DNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Extended transgene expression from a nonintegrating adenoviral vector containing retroviral elements. / Zheng, Changyu; Vitolo, Joseph M; Zhang, Weitian; Mineshiba, Fumi; Chiorini, John A.; Baum, Bruce J.

In: Molecular Therapy, Vol. 16, No. 6, 01.01.2008, p. 1089-1097.

Research output: Contribution to journalArticle

Zheng, Changyu ; Vitolo, Joseph M ; Zhang, Weitian ; Mineshiba, Fumi ; Chiorini, John A. ; Baum, Bruce J. / Extended transgene expression from a nonintegrating adenoviral vector containing retroviral elements. In: Molecular Therapy. 2008 ; Vol. 16, No. 6. pp. 1089-1097.
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