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An epi-allelic series of p53 hypomorphs created by stable RNAi produces distinct tumor phenotypes in vivo

Abstract

The application of RNA interference (RNAi) to mammalian systems has the potential to revolutionize genetics and produce novel therapies. Here we investigate whether RNAi applied to a well-characterized gene can stably suppress gene expression in hematopoietic stem cells and produce detectable phenotypes in mice. Deletion of the Trp53 tumor suppressor gene greatly accelerates Myc-induced lymphomagenesis, resulting in highly disseminated disease1,2. To determine whether RNAi suppression of Trp53 could produce a similar phenotype, we introduced several Trp53 short hairpin RNAs (shRNAs) into hematopoietic stem cells derived from Eμ-Myc transgenic mice, and monitored tumor onset and overall pathology in lethally irradiated recipients. Different Trp53 shRNAs produced distinct phenotypes in vivo, ranging from benign lymphoid hyperplasias to highly disseminated lymphomas that paralleled Trp53−/− lymphomagenesis in the Eμ-Myc mouse. In all cases, the severity and type of disease correlated with the extent to which specific shRNAs inhibited p53 activity. Therefore, RNAi can stably suppress gene expression in stem cells and reconstituted organs derived from those cells. In addition, intrinsic differences between individual shRNA expression vectors targeting the same gene can be used to create an 'epi-allelic series' for dissecting gene function in vivo.

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Figure 1: Analysis of Trp53 shRNA function in vitro.
Figure 2: Acceleration of Eμ-Myc–induced lymphomagenesis by Trp53 shRNAs.
Figure 3: Trp53 shRNAs produce distinct pathologies in vivo.
Figure 4: Trp53 shRNAs and chromosomal instability.
Figure 5: Trp53 shRNAs contribute to lymphomas and suppress mutations in endogenous Trp53.

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Acknowledgements

We thank J. Adams and T. Jacks for providing mice, and C. Rosenthal, M. E. Dudas, and M. S. Jiao for invaluable technical assistance. P.J.P. is an Arnold and Mabel Beckman Fellow of the Watson School of Biological Sciences. G.J.H. is a Rita Allen Foundation Scholar and is supported by an Innovator award from the U.S. Army Breast Cancer Research Program. This work was supported by US National Cancer Institute (NCI) postdoctoral training grants (M.T.H. and J.T.Z.), program project grants (C.C.C., G.J.H. and S.W.L.) from the NCI, and a generous gift from the Ann L. and Herbert J. Siegel Philanthropic Fund.

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Correspondence to Gregory J. Hannon or Scott W. Lowe.

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Hemann, M., Fridman, J., Zilfou, J. et al. An epi-allelic series of p53 hypomorphs created by stable RNAi produces distinct tumor phenotypes in vivo. Nat Genet 33, 396–400 (2003). https://doi.org/10.1038/ng1091

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