β-globin mRNA half-life in a tet-on (tetracycline-inducible promoter) HeLa cell

Value 13 hours
Organism Human Homo sapiens
Reference Audibert A, Weil D, Dautry F. In vivo kinetics of mRNA splicing and transport in mammalian cells. Mol Cell Biol. 2002 Oct22(19):6706-18. doi: 10.1128/MCB.22.19.6706-6718.2002 p.6711 left column top paragraph & p.6712 right column top linePubMed ID12215528
Method P.6714 left column top paragraph:"In this study [investigators] present a strategy to analyze the splicing and transport rates of pre-mRNA in living cells that relies on the use of regulated promoters. This approach associates the kinetic dimension of transcription regulation by tetracycline with the sensitivity of the RNase protection assay."
Comments p.6710 right column bottom paragraph:"In longer experiments, the steady state was still not reached after 60 h (data not shown). Because, once transcription is maximal, a plateau should be reached within a few transcript half-lives (e.g., 75% of maximal expression should be achieved in two half-lives), the absence of an approach to equilibrium could indicate either a very long half-life for β-globin mRNA or a continuous increase in transcription. To address this issue, [investigators] measured the β-globin mRNA half-life in these HeLa tet-on cells by washing off doxycycline. A Northern blot analysis and the quantification of the corresponding signals using β-actin as an internal standard are presented in Fig. 3C and indicate a β-globin mRNA half-life of 13 h. Thus, the failure to reach equilibrium after 53 h of induction (i.e., four mRNA half-lives) suggested that transcription kept increasing for at least 2 days after induction by doxycycline."
Entered by Uri M
ID 112187