AIM: To evaluate the effect of EffecteneTM lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA4-VEGI) gene on corneal neovascularization (CNV). METHODS: Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to induce CNV and divided into 4 random teams, ten per each team: team A: transfected by pcDNA4-VEGI gene mediated by EffecteneTM lipofectine transfection; team B: by plasmid pcDNA4; team C: by EffecteneTM, and team D: by normal saline. Length and area of CNV were observed under slit lamp every day after tran- sfection. Immunohistochemistry was performed to detect the expression of VEGI protein in corneas at day 3, 7, 14 and 21. RESULTS: 1) Average occurrence of CNV was 6.3 days in team A, 3.1 days in team B, 3.2 days in team C, and 3.2 days in team D. Difference was significant between A and other teams (P<0.01); 2) Length and average area of CNV in each period in team A was significantly different from those in team B, C and D (P<0.01); 3) VEGI expressions were observed in epithelium, stroma, endothelium and the cliff of CNV in team A at 3 days after transfection by immunohistochemical staining. None VEGI positive cells were found in the control teams (team B, C and D) all the time. CONCLUSION: EffecteneTM lipofectine transfection technique can effectively transfect pcDNA4-VEGI gene into rabbit cornea and the length and CNV areas can be inhibited by VEGI gene.