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Lentivirus vectors mediated eGFP transfected into rat ovary in vivo.

PURPOSE
To evaluate the optimum dosage and time-effect relationship of lentivirus vectors mediated enhanced green fluorescence protein gene (lenti-eGFP) transfection into the rat ovary in vivo.


MATERIALS AND METHODS
Lenti-eGFP was microinjected into rats' ovaries with different dosage (2 x 10(6) TU and 10 x 10(6) TU virosome respectively, n = 5 rats). The expression of eGFP was examined by the fluorescence microscope at injection at five days. The fluorescence intensity of different dose groups was calculated and determined the optimum dosage. The authors observed the expression of eGFP in ovaries and others tissues at days 5, 15, 30, 45, 60, and 75 after the rats' ovaries were microinjected with the optimum dosage of lenti-eGFP. Reserve transcription-polymerase chain reaction (RT-PCR) and RT-quantitative (q) PCR (RT-qPCR) were used to qualitative and quantitative analyze the expression of eGFP in different tissues and organs of transfected rats.


RESULTS
The expression of eGFP in both ovaries of every rat was seen at five days of transfection. Semi-quantitative assessment of green fluorescence for the two-dosage group was 0.2311 +/- 0.0203 and 0.2307 +/- 0.0199, respectively. There was no significant difference in both groups (p = 0.976). The expression of eGFP enhanced with transfection time prolongation and continued with 75 days of transfection (the fluorescence density in different time was 0.2307 +/- 0.0199, 0.3119 +/- 0.0213, 0.3462 +/- 0.0264, 0.3568 +/- 0.0127, 0.3496 +/- 0.0133, and 0.3513 +/- 0.0172, respectively). Furthermore, there were efficient and durable expressions of eGFP in other tissues and organs of rats. RT-PCR and RT-qPCR proved these results.


CONCLUSION
Lenti-eGFP may successfully transfect ovary tissues and other organs in vivo simultaneously, the expression of eGFP is highly efficient and durable.