Adenovirus-mediated gene transfer to keratinocytes--a review.
The introduction and expression of a foreign gene provide a powerful tool for investigating functions and regulation of a gene of interest; however, keratinocytes have a major drawback in that foreign genes are hardly transfected by conventional methods and stable transformants are most difficult to establish in normal keratinocytes with a limited short life span. To overcome these problems, we used an adenovirus vector, Ax, developed by Saito et al, which yields desired recombinant viruses at an efficiency about 100-fold that of conventional methods, and by which genes are expressed at a high level under the control of a composite CAG promoter. We established Ax vectors carrying various isoforms of protein kinase C (PKC). Using these vectors, we found that the eta and delta isoforms of PKC, but not the alpha and zeta isoforms, mediate terminal differentiation in normal human keratinocytes. These Ax-vectors are also applicable to organ culture of mouse embryos. Advantages and disadvantages of adenovirus vectors and their use for keratinocyte biology are reviewed.
Kuroki
T
T
The Institute of Molecular Oncology, Showa University, Hatanodai, Tokyo, Japan.
Kashiwagi
M
M
Ishino
K
K
Huh
N
N
Ohba
M
M
D000256:Adenoviridae; D000818:Animals; D002478:Cells, Cultured; D016193:G1 Phase; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D015603:Keratinocytes / Q000378:metabolism* / Q000821:virology; D011493:Protein Kinase C / Q000235:genetics
D007527:Isoenzymes; D011493:Protein Kinase C