Simultaneous detection of amplicon and HSV-1 helper encoded proteins reveals that neurons and astrocytoma cells do express amplicon-borne transgenes in the absence of synthesis of virus immediate early proteins.
HSV-1 amplicon vectors were used to express either a cytoplasmic (beta-galactosidase) or a membrane targeted protein (TIMP-Thy1) in primary neuronal cultures, and a human astrocytoma cell line. Whereas some cells became infected by vector particles alone others were simultaneously infected by both vector and helper particles. Our results show that IEHCMV and HSV-1 IE3 promoters are able to direct transgene expression in these cells in the absence of synthesis of helper virus transacting proteins, and stress the need of monitoring expression from both partners of an amplicon population, in order to differentiate transgene expression in cells singly infected with amplicon particles, from those infected by both amplicon and helper particles.
Lowenstein
P R
PR
Laboratory of Cellular and Molecular Neurobiology, School of Molecular and Biomedical Sciences, University of Wales College of Cardiff, UK.
Fournel
S
S
Bain
D
D
Tomasec
P
P
Clissold
P M
PM
Castro
M G
MG
Epstein
A L
AL
D001253:Astrocytes / Q000378:metabolism; D001254:Astrocytoma / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000235:genetics; D009474:Neurons / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
D017874:Immediate-Early Proteins; D014764:Viral Proteins