A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver

Kirk J. Wangensteen, Andrew Wilber, Vincent W. Keng, Zhiying He, Ilze Matise, Laura Wangensteen, Corey M. Carson, Yixin Chen, Clifford J Steer, R S Mc Ivor, David A Largaespada, Xin Wang, Stephen C. Ekker

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Current techniques for the alteration of gene expression in the liver have a number of limitations, including the lack of stable somatic gene transfer and the technical challenges of germline transgenesis. Rapid and stable genetic engineering of the liver would allow systematic, in vivo testing of contributions by many genes to disease. After fumaryl acetoacetate hydrolase (Fah) gene transfer to hepatocytes, selective repopulation of the liver occurs in FAH-deficient mice. This genetic correction is readily mediated with transposons. Using this approach, we show that genes with biological utility can be linked to a selectable Fah transposon cassette. First, net conversion of Fah-/- liver tissue to transgenic tissue, and its outgrowth, was monitored by bioluminescence in vivo from a luciferase gene linked to the FAH gene. Second, coexpressed short hairpin RNAs (shRNAs) stably reduced target gene expression, indicating the potential for loss-of-function assays. Third, a mutant allele of human α1-antitrypsin (hAAT) was linked to Fah and resulted in protein inclusions within hepatocytes, which are the histopathological hallmark of hAAT deficiency disorder. Finally, oncogenes linked to Fah resulted in transformation of transduced hepatocytes. Conclusion: Coexpression with FAH is an effective technique for lifelong expression of transgenes in adult hepatocytes with applicability to a wide variety of genetic studies in the liver.

Original languageEnglish (US)
Pages (from-to)1714-1724
Number of pages11
JournalHepatology
Volume47
Issue number5
DOIs
StatePublished - May 1 2008

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Hydrolases
Hepatocytes
Liver
Genes
Gene Expression
Gene Transfer Techniques
Genetic Engineering
Luciferases
Transgenes
Oncogenes
Small Interfering RNA
Alleles
acetoacetic acid
Proteins

Cite this

Wangensteen, K. J., Wilber, A., Keng, V. W., He, Z., Matise, I., Wangensteen, L., ... Ekker, S. C. (2008). A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver. Hepatology, 47(5), 1714-1724. https://doi.org/10.1002/hep.22195

A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver. / Wangensteen, Kirk J.; Wilber, Andrew; Keng, Vincent W.; He, Zhiying; Matise, Ilze; Wangensteen, Laura; Carson, Corey M.; Chen, Yixin; Steer, Clifford J; Mc Ivor, R S; Largaespada, David A; Wang, Xin; Ekker, Stephen C.

In: Hepatology, Vol. 47, No. 5, 01.05.2008, p. 1714-1724.

Research output: Contribution to journalArticle

Wangensteen, KJ, Wilber, A, Keng, VW, He, Z, Matise, I, Wangensteen, L, Carson, CM, Chen, Y, Steer, CJ, Mc Ivor, RS, Largaespada, DA, Wang, X & Ekker, SC 2008, 'A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver', Hepatology, vol. 47, no. 5, pp. 1714-1724. https://doi.org/10.1002/hep.22195
Wangensteen KJ, Wilber A, Keng VW, He Z, Matise I, Wangensteen L et al. A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver. Hepatology. 2008 May 1;47(5):1714-1724. https://doi.org/10.1002/hep.22195
Wangensteen, Kirk J. ; Wilber, Andrew ; Keng, Vincent W. ; He, Zhiying ; Matise, Ilze ; Wangensteen, Laura ; Carson, Corey M. ; Chen, Yixin ; Steer, Clifford J ; Mc Ivor, R S ; Largaespada, David A ; Wang, Xin ; Ekker, Stephen C. / A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver. In: Hepatology. 2008 ; Vol. 47, No. 5. pp. 1714-1724.
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