A Flexible Approach to Studying Post-Transcriptional Gene Regulation in Stably Transfected Mammalian Cells

作者：Nichols Ralph C^*; Botson John; Wang Xiao Wei; Hamilton B JoNell; Collins Jane E; Uribe Victoria; Brooks Seth A; Zan Moe; Rigby William F C

来源：Molecular Biotechnology, 2011, 48(3): 210-217.

DOI：10.1007/s12033-010-9360-8

• ARE
• TNF
• mRNA turnover
• Translation
• 3 '-UTR

The study of post-transcriptional regulation is constrained by the technical limitations associated with both transient and stable transfection of chimeric reporter plasmids examining the activity of 3'-UTR cis-acting elements. We report the adaptation of a commercially available system that enables consistent stable integration of chimeric reporter cDNA into a single genomic site in which transcription is induced by tetracycline. Using this system, we demonstrate the tight control afforded by this system and its suitability in mapping the regulatory function of defined cis-acting elements in the human TNF 3'-UTR, as well as the distinct effects of serum starvation on transiently transfected and stably integrated chimeric reporter genes.

• 出版日期2011-7