A 7.1 kbp beta-myosin heavy chain promoter, efficient for green fluorescent protein expression, probably induces lethality when overexpressing a mutated transforming growth factor-beta type II receptor in transgenic mice.

Autor: Sverine Allegra, Lamia Bouazza, Claire Benetollo, Jacques Yuan Li, Dominique Langlois
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Zdroj: Transgenic Research; Feb2005, Vol. 14 Issue 1, p69-80, 12p
Abstrakt: Abstract The roles of transforming growth factor-beta (TGF) in heart or skeletal muscle development and physiology are still the subject of controversies. Our aim was to block, in transgenic mice, the TGF signalling pathway by a dominant negative mutant of the TGF type II receptor fused to the enhanced green fluorescent protein (TRII-KR-EGFP) under the control of a 7.1 kbp mouse beta-myosin heavy chain (MHC) promoter to investigate the roles of TGF in the heart and slow skeletal muscles. First, we generated two transgenic lines overexpressing EGFP under the control of the 7.1 kbp MHC promoter. In embryos, EGFP was detectable as early as 7.5 days post coitum. In embryos, newborns and adults, EGFP was expressed mainly in the cardiac ventricles and in slow skeletal muscles. EGFP expression was intense in the bladder but weak in the intestines. In contrast to the endogenous MHC promoter, the activity of the 7.1 kbp MHC promoter in the transgene was not repressed after birth and remained high in adult transgenic mice. We obtained two founders with the transgene comprising the TRII-KR-EGFP sequence under the control of the 7.1 kbp MHC promoter. These founders were generated at a very low frequency and expressed barely detectable levels of TRII-KR-EGFP mRNA. Our failure to obtain transgenic lines overexpressing the dominant negative receptor suggests that the blocking of the TGF signalling pathway in the heart and slow skeletal muscles could be embryonically lethal. To conclude, the 7.1 kbp MHC promoter directs high levels of transgene expression in the cardiac ventricles and in slow skeletal muscles of the mouse. Analysis of the consequences of the blocking of the TGF signalling pathway in the heart will require the use of tissue specific means of conditional gene invalidation. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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