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A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver
Author(s) -
Wangensteen Kirk J.,
Wilber Andrew,
Keng Vincent W.,
He Zhiying,
Matise Ilze,
Wangensteen Laura,
Carson Corey M.,
Chen Yixin,
Steer Clifford J.,
McIvor R. Scott,
Largaespada David A.,
Wang Xin,
Ekker Stephen C.
Publication year - 2008
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.22195
Subject(s) - biology , transgene , gene , reporter gene , selectable marker , gene expression , somatic cell , microbiology and biotechnology , gene knockin , genetics
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. (H EPATOLOGY 2008;47:1714–1724.)