By R. Stone, J. Stone

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Extra info for Atlas of Skeletal Muscles

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They then translated these mutations to the tTA transactivator to generate the socalled reverse tTA (rtTA), because it exhibits the reverse behavior of tTA: rtTA can bind to the TRE and activate downstream genes only in the presence of tetracycline or its derivatives (Fig. 1). In summary, addition of tetracycline blocks transcription in the Tet-Off system, while it activates transcription in the Tet-On system. These systems have been extensively used in cell culture and stable cell lines containing either the tTA or rtTA regulatory protein have been established.

References Das, R. , Van Hateren, N. , Howell, G. , Farrell, E. , Bangs, F. , Porteous, V. , Manning, E. , McGrew, M. , Sacco, M. , Halley, P. , Sang, H. , Storey, K. , Nair, V. , Wilson, S. A. (2006) A robust system for RNA interference in the chicken using a modified microRNA operon. Dev Biol 294, 554–563. , Nakamura, H. (1999) Role of Pax-5 in the regulation of a mid-hindbrainorganizer’s activity. Dev Growth Differ 41, 59–72. , Hamilton, H. (1951) A series of normal stages in the development of the chick embryo.

Plasmid DNA is extracted using an Endotoxin Free Qiagen kit. 2 Electroporation Electroporations are performed using established procedures. For an extensive description, the reader can refer to the different chapters of this book. In the case of studying mRNA stability in the neural tube, electroporations are performed as follow: 4 Temporal Control of Gene Expression by Combining Electroporation 29 A window is opened in the shell of 2-day-old embryos (12 to 18 somites). 5% Fast Green in PBS is injected into the lumen of the neural tube.

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