Stable levels of long-term transgene expression driven by the latency-associated transcript promoter in a herpes simplex virus type 1 vector.
Previous gene transfer studies of the herpes simplex virus type 1 (HSV-1) using the latency-associated transcript (LAT) promoter have reported a decrease in transgene expression in the brain over time, but the extent of this decrease has not been measured and it is unknown if expression eventually stabilizes. We examined LAT promoter-mediated transgene expression in the mouse brain for 1 year following intracranial injection with a HSV-1 vector expressing human beta-glucuronidase (GUSB). The vector genome copy number remained stable from 2 to 52 weeks. Quantitative reverse transcriptase PCR detected a peak of LAT intron expression at 2 weeks (corresponding to the end of the acute phase of viral infection), followed by stable expression during latency (13-52 weeks). The number of GUSB-positive cells also had a peak in the acute phase and then was stable during latency (13-52 weeks). GUSB enzymatic activity was maintained at 11% of normal at 6 and 12 months, indicating that the LAT promoter is capable of driving stable transgene expression in the brain.
Berges
B K
BK
Department of Microbiology, School of Medicine, 319 Johnson Pavilion, University of Pennsylvania, Philadelphia, PA 19104, USA.
Wolfe
J H
JH
Fraser
N W
NW
D000818:Animals; D001921:Brain / Q000378:metabolism; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D007438:Introns; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018139:Simplexvirus / Q000235:genetics*; D013997:Time Factors; D019076:Transgenes; D014709:Vero Cells
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004247:DNA; D005966:Glucuronidase