IMR Press / CEOG / Volume 40 / Issue 1 / pii/1630388034335-1640279937

Clinical and Experimental Obstetrics & Gynecology (CEOG) is published by IMR Press from Volume 46 Issue 1 (2019). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with S.O.G.

Open Access Origianal Research
Lentivirus vectors mediated eGFP transfected into rat ovary in vivo
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1 Department of Gynecology and Obstetrics of the First Affiliated Hospital of Chongqing Medical University, Chongqing City
2 Department of Gynecology and Obstetrics, Suining Central Hospital, Suining City
3 Department of Gynecology and Obstetrics, Zhongshan Hospital, Wuhan City
4 Department of Respiratory of Shandong Province Chest Hospital, Jinan City (China)
Clin. Exp. Obstet. Gynecol. 2013, 40(1), 101–105;
Published: 10 March 2013
Abstract

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 × 106 TU and 10 × 106 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.
Keywords
Lentivirus vectors
Enhanced green fluorescence protein
Ovary
In vivo
Animal experiment
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