Identification of multiple sites suitable for insertion of foreign genes in herpes simplex virus genomes.
Information on sites in HSV genomes at which foreign gene(s) can be inserted without disrupting viral genes or affecting properties of the parental virus are important for basic research on HSV and development of HSV-based vectors for human therapy. The intergenic region between HSV-1 UL3 and UL4 genes has been reported to satisfy the requirements for such an insertion site. The UL3 and UL4 genes are oriented toward the intergenic region and, therefore, insertion of a foreign gene(s) into the region between the UL3 and UL4 polyadenylation signals should not disrupt any viral genes or transcriptional units. HSV-1 and HSV-2 each have more than 10 additional regions structurally similar to the intergenic region between UL3 and UL4. In the studies reported here, it has been demonstrated that insertion of a reporter gene expression cassette into several of the HSV-1 and HSV-2 intergenic regions has no effect on viral growth in cell culture or virulence in mice, suggesting that these multiple intergenic regions may be suitable HSV sites for insertion of foreign genes.
Morimoto
Tomomi
T
Division of Viral Infection, Department of Infectious Disease Control, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639.
Arii
Jun
J
Akashi
Hiroomi
H
Kawaguchi
Yasushi
Y
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D021901:DNA, Intergenic; D005260:Female; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006561:Herpes Simplex / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D016254:Mutagenesis, Insertional; D011817:Rabbits; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics
D021901:DNA, Intergenic; C121640:UL3 protein, Human herpesvirus 1; C082590:UL4 protein, Human cytomegalovirus; D014759:Viral Envelope Proteins; D014764:Viral Proteins