关键词: 慢病毒, 曲细精管, 小鼠, 转基因

Abstract: Objective To investigate the practicability of lentiviral vector-based sperm-mediated gene transfer in production of transgenic animals.Methods According to the titer of lentiviral vector (LV) administrated, 4～5 week-old mice were divided into two groups: high-titer (2 mice) and low-titer (6 mice) groups, into which lentiviral vectors carrying green fluorescent protein (LV-GFP) were injected through the seminiferous tubules at the concentration of 20μL/testis, and the administrated titer of LV was 1×109 and 2×10 7 TU/mL for the two groups respectively. Sampling was conducted at the 4th and 8th week (high-titer group), and the 5th, 13th and 17th week (low-titer group) after injection. Transgenic mice were examined by PCR, and the expression of GFP was detected by fluorescence microscopy and immunohistochemistry.Results (1) positive rate of GFP gene in mice testis was 62.5%; (2) weak expression of GFP was only observed in high-titer group, and mainly distributed in the based-membrane of seminiferous tubules and the interspaces between tubules; (3) injection into seminiferous tubules caused some damage, but did not affect the reproductive capacity of mice. Conclusion Lentiviral vectors can successfully transfect mice testis in SMGT and the expression of GFP gene depends on the titer of LV administrated.

Key words: Lentiviral vector, Sperm-mediated gene transfer, Sminiferous tubules, Mice