Establishment of SV40-tsA58 transgenic rats as a source of conditionally immortalized cell lines.
To isolate a variety of rat cell lines with differentiated functions, we established transgenic rat lines expressing the temperature-sensitive large T-antigen of simian virus 40 (SV40) tsA58 mutant under the control of the SV40 large T-antigen itself. We microinjected the DNA into 564 eggs of Wistar rat and 23 independent transgenic candidates were obtained. Ten pups died before weaning and eight transgenic rats could not transmit the transgene to the progeny. Finally, five lines of the transgenic rat were established. Although one line (#1511-6) had low reproductivity, the other four lines reproduced normally. Three out of the four lines (#1507-2, #1509-7, #1519-8) appeared normal but the other line had tumors in the brain and subcutaneous tissue at 3 weeks of age (#1511-6), and in the kidneys and subcutaneous tissue at 18 to 19-weeks of age (#1507-5). Fibroblast cells prepared from transgenic fetuses of lines #1507-5 and #1519-8 expressed the transgene and exhibited temperature-dependent growth. Both of the lines (#1507-5 and #1519-8) were successfully generated to be homozygous by sibling mating of transgenic offspring. These transgenic rat lines have bred through many generations and have been established to be a ready source of novel conditionally immortalized cell lines.
Takahashi
R
R
YS New Technology Institute Inc., Tochigi, Japan.
Hirabayashi
M
M
Yanai
N
N
Obinata
M
M
Ueda
M
M
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D005814:Genes, Viral / Q000502:physiology; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D010063:Ovum / Q000821:virology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D013539:Simian virus 40 / Q000235:genetics*
D000952:Antigens, Polyomavirus Transforming