[Preparation of green fluorescent protein retrovirus and its application in mediating gene transfer into retinal pigment epithelial cells].
OBJECTIVE
To determine whether human retinal pigment epithelial (RPE) cells can be infected by retrovirus and modified by retrovirus-mediated gene transfer.


METHODS
Recombination retroviral vector pLNCX-GFP (green fluorescent protein, GFP) was generated by inserting 780 bp GFP cDNA fragment into the MCS site of pLNCX. pLNCX-GFP was transfected into ecotropic packaging cell line PhiX-Eco and amphotropic packaging cell line PhiX-Ampho and PA317. Retroviral titer was tested by counting GFP expression of NIH3T3 cells. Then RPE cells were infected by using GFP retrovirus-containing supernatant.


RESULTS
GFP was expressed and retrovirus was produced upon pLNCX-GFP being transfected into packaging cell line. The GFP retrovirus was able to infect primary cultured human RPE cells and immortalized RPE cell line.


CONCLUSIONS
The retrovirus can introduce a foreign gene into RPE cells efficiently, thereby it can be used as an important tool to deliver gene into RPE for therapy of fundus diseases.
Huang
Q
Q
Central Laboratory, The First People's Hospital of Shanghai, Shanghai 200080, China.
Xu
P
P
Liu
W
W
Wang
F
F
Gu
Q
Q
Tian
S
S
Fan
Y
Y
Xig
K
K
Chen
X
X
Li
C
C
D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D010857:Pigment Epithelium of Eye / Q000166:cytology*; D012190:Retroviridae / Q000235:genetics*
D049452:Green Fluorescent Proteins