December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
Production of a Lentiviral Vector Containing Tissue Inhibitor of Metalloproteinase 1, an Inhibitor of Angiogenesis
Author Affiliations & Notes
  • AC Westfall
    Ophthalmology Casey Eye-Oregon Hlth Sci Univ Portland OR
  • Footnotes
    Commercial Relationships   A.C. Westfall, None.
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1762. doi:
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      AC Westfall; Production of a Lentiviral Vector Containing Tissue Inhibitor of Metalloproteinase 1, an Inhibitor of Angiogenesis . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1762.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract: : Purpose: Protein tissue inhibitor of metalloproteinase-1 (TIMP-1) is capable of inhibiting tumor growth, invasion, metastasis and also inhibits angiogenesis. We sought to develop a potentially therapeutic reagent, able to efficiently deliver an expressible TIMP-1 gene. Methods: TIMP-1 cDNA was cloned into the lentiviral vector pHR'-IRES-eGFP under the control of the CMV promoter. Replication-defective TIMP-1 lentivirus was prepared by co-transfection of three plasmids, pHR'CMV-TIMP-1-IRES-eGFP, pCMV Δ;R8.91 and pMD.G, into 293T cells. A 4 x 4mm nylon mesh soaked with the recombinant TIMP-1 lentivirus was inserted into a corneal stromal pocket in one eye of New Zealand White rabbits, contralateral eyes were evaluated as nontransduced eyes. An additional control included corneal pockets containing mesh soaked with pHR'-IRES-eGFP virus (TIMP-1 deficient). Assay of the development of corneal neovascularization is underway Results: A 624 bp cDNA encoding a 207 aa protein for human TIMP-1, a zinc-binding peptidase, was successfully cloned into a pHR'CMV-IRES-eGFP plasmid. Recombinant lentiviral pHR'CMV-TIMP-1-IRES-eGFP was produced. Conclusions: TIMP-1 is involved in inhibiting angiogenesis. The construction of a lentiviral vector that can efficiently introduce TIMP-1 into human cells is a useful tool for studying angiogenesis in in vitro and in vivo models. Supported by the Clayton Foundation for Research and Research to Prevent Blindness

Keywords: 419 gene transfer/gene therapy • 483 neovascularization 

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