Establishment and identification of GFP transgenic mice model mediated by lentiviral vector

Objective To establish a technology platform of transgenic animals mediated by lentiviral vector through a practice of preparing the transgenic mice by lentiviral vector carrying green fluorescent protein(GFP).Methods 293FT cells were transfected by a mixture of leniviral vector FUGW and viraPowerTM lentiviral packaging mix(1∶2) using LipofectamineTM 2000.After undergoing concentration,the titer of packaged lentivirus was tested in 293T cells.The transgenic mice were reproduced by injecting the lentiviral vectors into the perivitelline space.Integration of exogenous gene in transgenic mice was confirmed by PCR detection,and the expression of exogenous gene in transgenic mice was identified by fluorescence stereomicroscope.Results Twenty-four hours later,GFP expression was detected in 293FT cells.A great amount of green fluorescence can be detected under fluorescence inverted microscope after 72 hours.The transfection efficiency of 293FT cells reached to higher than 95%.The virus titer of packaged lentiviral vector was 106U/ml,and was 108U/ml after concentration.In F0 generation,GFP could be detected in 70.3%(26/37)transgenic mice by PCR.Under fluorescence stereomicroscope,GFP expressed in 65.2%(15/23) and 55.0%(11/20) of F0 and F1 generation,respectively,and the level was comparable.The GFP expression was also widely observed in both F0 and F1 generations.Conclusions The transgenic mice model carrying GFP is produced and the GFP gene can be transmitted through germline cells.The technology platform of transgenic mice mediated by lentiviral vector has been successfully established.