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Other transgenic mutation assays: Development of a novel mouse tk+/− embryonic stem cell line for use in mutagenicity studies

✍ Scribed by Vasily N. Dobrovolsky; Daniel A. Casciano; Robert H. Heflich


Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
663 KB
Volume
28
Category
Article
ISSN
0893-6692

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✦ Synopsis


A tk+/mouse embryonic stem (ES) cell line, designated 1G2, has been created in which one allele of the thymidine kinase (tk) gene was inactivated by targeted homologous recombination. This line is an analog of the mouse lymphoma tk+/-L5 178Y cell line, which is used widely to assess the mutagenicity of chemical agents. Treatment of 1 G 2 cells with the alkylating agent Nethyl-Nnitrosourea (ENUJ resulted in a doserelated increase in triflu-orothymidine-resistant colonies. Mutant frequencies of 152 and 296 per 1 O6 cells were determined for 0.1 and 0.3 mg/ml doses of ENU, compared with a spontaneous mutant frequency of 1 5 per 106 cells. The data indicate that tk+/-1 G 2 ES cells may be useful for the creation of a transgenic mouse model for assessing in vivo mutation using an endogenous autosomal gene. o 1996 WiIey-Liss, Inc.