Characterisation of eGFP-transgenic BALB/c mouse strain established by lentiviral transgenesis.
Lentiviral technology is a powerful tool for the creation of stable transgenic animals. However, uncertainties have remained whether constitutive promoters resist long-term silencing. We used concentrated HIV-1 based lentiviral vectors to create stable transgenic BALB/c mice by perivitelline injection. In our vectors eGFP expression was driven by the human EF1alpha promoter. The established transgenic animals were analyzed for eGFP expression by in vivo fluorescence imaging, PCR, histology and flow-cytometry. eGFP expression showed even distribution without mosaicism; however, tissue-dependent differences of eGFP expression were observed. Up to the sixth generation only one newborn showed eGFP inactivation. eGFP + transgenic bone marrow cells efficiently provided long-term haemopoietic repopulation in radiation chimeras, regenerating all bone marrow-derived lineages with eGFP + cells with distinct eGFP expression profiles. The established eGFP + BALB/c mouse strain is expected to be extremely useful in various immunological experiments.
Kvell
Krisztián
K
Department of Immunology and Biotechnology, University of Pecs, Pecs, Hungary.
Czömpöly
Tamás
T
Hiripi
László
L
Balogh
Péter
P
Kóbor
József
J
Bodrogi
Lilla
L
Pongrácz
Judit E
JE
Ritchie
William A
WA
Bosze
Zsuzsanna
Z
D000818:Animals; D000831:Animals, Newborn; D004526:Efficiency; D005260:Female; D020868:Gene Silencing / Q000502:physiology; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics*; D040582:Inheritance Patterns; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics
D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins