[Preparation of green fluorescent protein retrovirus and its application in mediating gene transfer into retinal pigment epithelial cells].


Journal Article

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. CONCLUSION: 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.

Full Text

Duke Authors

Cited Authors

  • Huang, Q; Xu, P; Liu, W; Wang, F; Gu, Q; Tian, S; Fan, Y; Xig, K; Chen, X; Li, C

Published Date

  • July 2001

Published In

Volume / Issue

  • 37 / 4

Start / End Page

  • 248 - 251

PubMed ID

  • 11864429

Pubmed Central ID

  • 11864429

International Standard Serial Number (ISSN)

  • 0412-4081


  • chi

Conference Location

  • China