Arch Virol (2004) 149: 1653–1662
Selection of genetic inhibitors of rabies virus
W. H. Wunner
, C. Pallatroni
, and P. J. Curtis
The Wistar Institute, Philadelphia, Pennsylvania, U.S.A.
Department of Microbiology and Immunology, Jefferson Alumni Hall,
Thomas Jefferson University, Philadelphia, Pennsylvania, U.S.A.
Received February 12, 2003; accepted January 15, 2004
Published online April 15, 2004
Summary. A cDNA library of short random fragments derived from four of the
ﬁve genes of the rabies virus genome has been used to isolate genetic suppressor
elements (GSEs) expressed intracellularly that inhibit rabies virus replication. Two
nucleotide fragments, one from the rabies virus nucleocapsid protein (N) gene
and the other from the phosphoprotein (P) gene, have been identiﬁed as inhibitors
of rabies virus replication in cell culture. The N cDNA fragment is expressed
in sense-orientation and could produce a dominant negative protein affecting
virus replication. The P cDNA fragment is expressed in the inhibitory antisense
direction. Inhibition of rabies virus replication was detected in cell culture using
an ELISA for detection of rabies virus glycoprotein expression on the cell surface
and immunoﬂuorescence for detection of intracellular rabies virus N expression.
Both the sense and antisense GSEs, because of their targeted inhibition of rabies
virus replication, have possible uses in rational design of antiviral compounds for
treatment of rabies. This approach could be applied to any virus, particularly to
those that lyse their target host cell.
Rabies is a fatal human disease that is prevented only by preexposure (prophylatic)
immunization (PEI) with vaccine or postexposure treatment (PET) with vac-
cine for active immunization plus antirabies immunoglobulin of horse or human
(HRIG) origin for passive immunization. Rabies in humans remains a serious
public health problem in many countries, including the United States, where
rabies virus transmission among wildlife or domestic animal reservoirs continues
to threaten humans exposed to rabies virus infected animals . Rabies vaccines
have been developed for PEI and PET of humans, but at times these, particularly