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Nucleic Acids Research, 1995, Vol. 23, No. 9 1495-1501
© 1995

MOLECULAR BIOLOGY

A sindbis virus mRNA polynucleotide vector achieves prolonged and high level heterologous gene expression in vivo

F. W. Johanning1, R. M. Conry2, A. F. LoBuglio3, M. Wright1, L. A. Sumerel2, M. J. Pike2 and D. T. Curiel1,3,*

1Gene Therapy Program Birmingham, AL 35294, USA 2Division of Hematology/Oncology Birmingham, AL 35294, USA 3Comprehensive Cancer Center, University of Alabama at Birmingham Birmingham, AL 35294, USA

*To whom correspondence should be addressed

Received February 8, 1995. Revised March 23, 1995. Accepted March 23, 1995.

The direct Intramuscular delivery of naked plasmld DNA has been demonstrated to allow expression of encoded heterologous genes in the target myocytes. The method has been employed to elicit Immunization based upon delivery of antigen encoding plasmld DNA. For application In the context of achieving anti-tumor immunization against antigenic transforming oncoproteins, delivery of plasmld DNAs encoding these molecules would create significant potential safety hazards. As an alternative to DNA polynucleotide vectors, we explored the utility of mRNA vehicles for Inducing foreign gene expression in muscle cells In vivo. Synthetic reporter-gene encoding mRNA transcripts were derived for this analysis. The sindbis virus vector was also used to derive luciferase mRNA transcripts which possessed self-replication capacity. In these studies, It could be shown that the replicative vector was capable of directing significantly elevated levels of reporter gene expression In myocytes compared to a non-replicatlve mRNA species. In addition, the replicative species was capable of achieving significantly prolonged levels of In vivo gene expression compared to non-replicatlve mRNA. Both of these characteristics will make replicative mRNA vectors of utility for polynucleotide-based immunization protocols.


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