Differential neuropeptide Y gene expression in post-mitotic versus dividing neuroblastoma cells driven by an adeno-associated virus vector
Details
Serval ID
serval:BIB_8A1EE74A5EEB
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Differential neuropeptide Y gene expression in post-mitotic versus dividing neuroblastoma cells driven by an adeno-associated virus vector
Journal
Brain Research. Molecular Brain Research
ISSN
0169-328X (Print)
Publication state
Published
Issued date
07/1994
Volume
24
Number
1-4
Pages
27-33
Notes
Comparative Study
Journal Article --- Old month value: Jul
Journal Article --- Old month value: Jul
Abstract
The ability to express exogenous mammalian genes stably in post-mitotic cells such as neurons remains an important goal for those attempting to modulate neurotransmission through gene delivery. We therefore investigated how differentiation to a post-mitotic state affected the expression of an exogenous gene encoding for neuropeptide Y (NPY) following transfection with an adeno-associated virus (AAV) derived vector. This vector (pJDT95npy) was constructed with rat NPY cDNA (551 bp) inserted downstream from the indigenous AAV p5, p19 and p40 promoters to characterize their relative abilities to drive NPY mRNA expression. Transfection of dividing neuroblastoma CHP126 cells with pJDT95npy resulted in the differential expression of chimeric NPY mRNAs derived from each promoter. P40-driven species became dominant after 1 month post-transfection. Vector integration into chromosomal DNA was demonstrated by Southern blot analyses, indicating at least some region-selective integration. In dividing cell extracts, only a low level of pro-NPY immunoreactivity and no mature NPY immunoreactivity was recovered. However, after differentiation of the pJDT95npy-transfected CHP 126 cells to a post-mitotic state, significant levels of pro-NPY and mature NPY were recovered in the cells and media. Differentiation also had a time-dependent effect on mRNA expression: a spike of p5 driven expression on day 3 was followed predominantly by p40-driven expression on day 5. This study indicates that AAV-derived vectors using the p40 promoter may be used to express genes in post-mitotic cells such as neurons.
Keywords
Animals
Blotting, Northern
Blotting, Southern
Bucladesine/pharmacology
Cell Differentiation/drug effects
Cell Division
Cell Line
Dependovirus/*genetics
*Gene Expression
Genetic Vectors
Humans
Mitosis
Neuroblastoma/*metabolism
Neuropeptide Y/*biosynthesis
Plasmids
RNA, Messenger/analysis/biosynthesis
Rats
Recombinant Proteins/biosynthesis
Restriction Mapping
Transfection
Tumor Cells, Cultured
Pubmed
Create date
25/01/2008 10:55
Last modification date
20/08/2019 14:49