OBJECTIVE To evaluate the effect of Effectene™ lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA(4)-VEGI) gene on corneal neovascularization (CNV). METHODS Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to establish the animal model and divided into 4 random group, ten per each group: group A: transfected by pcDNA(4)-VEGI gene mediated by Effectene™ lipofectine transfection, group B: by Plasmid pcDNA(4), group C: by Effectene™, and group D: by normal saline. Length and area of CNV were measured under slit lamp every day after transfection, immunohistochemistry was used to detected the expression of VEGI protein in cornea at 3, 7, 14 and 21 days. RESULTS Average occurrence of CNV in the pcDNA(4)-VEGI gene transfected group (group A) was 6.3 days, in plasmid pcDNA(4) control group (group B) was 3.1 days, in Effectene™ lipofectine control group (group C) was 3.2 days, in normal saline control group (group D) was 3.2 days. Differences between groups A and B, C, D were statistically significant (P<0.01), while differences in groups B, C and D were meaningless (P>0.05). Lenth and average area of CNV in each period in group A was meaningful different from that in groups B, C, and D (P<0.01), while differences in group B, C and D were meaningless (P>0.05). Immunohistochemistry result: VEGI positive cells could be seen in epithelium, stroma, endothelium and the cliff of CNV in group A at 3 days after transfection. VEGI cells changed with the decrease of CNV. None positive cells were in the control groups (groups B, C and D) all the time. CONCLUSIONS Effectene™ lipofectine transfection technique can be effectively used in transfecting pcDNA(4)-VEGI gene into rabbit cornea and the lenth and areas of CNV can be inhibited by VEGI gene.