Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus.
The introduction of high molecular weight DNA into mammalian cells is useful for gene expression studies. However, current transfection strategies are inefficient, necessitating propagation of stable DNA transformants prior to analysis of gene expression. Here we demonstrate that transient lipid-mediated DNA transfection can be used to assess gene expression from yeast artificial chromosomes (YACs) containing the 230 kb cystic fibrosis transmembrane conductance regulator gene ( CFTR ) and Escherichia coli lacZ . We also show that psoralen-UV inactivated adenovirus significantly enhances transfection efficiency. The ability to deliver high molecular weight DNA using lipid-mediated transfection should expedite the analysis of large human genes contained within artificial chromosome vectors.
Chen
M
M
Vector Development Section, Genetics and Molecular Biology Branch and Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Compton
S T
ST
Coviello
V F
VF
Green
E D
ED
Ashlock
M A
MA
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002411:Cation Exchange Resins; D002460:Cell Line; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005363:Ficusin; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007763:Lac Operon / Q000235:genetics; D008055:Lipids; D017319:Photosensitizing Agents; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection / Q000379:methods*; D014466:Ultraviolet Rays
C505032:CFTR protein, human; D002411:Cation Exchange Resins; D008055:Lipids; C086724:Lipofectamine; D017319:Photosensitizing Agents; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005363:Ficusin