Abstract
A high saturated fat diet induces fatty liver in mice. The fatty liver is triggered initially by an increase in PPARγ2 protein in the liver, which leads to increased expression of lipogenic genes. Inactivation of PPARγ2 may improve fatty liver induced by a high saturated fat diet. RNA interference of PPARγ2 should be a powerful tool to elucidate the role of PPARγ2 in the development of fatty liver. Here, we describe our method for constructing the adenovirus vector containing short hairpin RNA targeted for PPARγ2. Adenovirus vector-mediated RNA interference for PPARγ2 should be useful for clarifying the biological role of the PPARγ2 pathway in various tissues and for therapeutic application in a variety of diseases.
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References
Graham FL, Smiley J, Russell WC, Nairn R (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74
Hammond SM, Caudy AA, Hannon GJ (2001) Post-transcriptional gene silencing by double-stranded RNA. Nat Rev Genet 2:110–119
Nykanen A, Haley B, Zamore PD (2001) ATP requirements and small interfering RNA structure in the RNA interference pathway. Cell 107:309–321
Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553
Paddison PJ, Caudy AA, Bernstein E, Hannon GJ, Conklin DS (2002) Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev 16:948–958
Paul CP, Good PD, Winer I, Engelke DR (2002) Effective expression of small interfering RNA in human cells. Nat Biotechnol 20:505–508
Berkner KL, Sharp PA (1983) Generation of adenovirus by transfection of plasmids. Nucleic Acids Res 11:6003–6020
Miyake S, Makimura M, Kanegae Y, Harada S, Sato Y, Takamori K, Tokuda C, Saito I (1996) Efficient generation of recombinant adenoviruses using adenovirus DNA-terminal protein complex and a cosmid bearing the full-length virus genome. Proc Natl Acad Sci USA 93:1320–1324
Fechner H, Haack A, Wang H, Wang X, Eizema K, Pauschinger M, Schoemaker R, Veghel R, Houtsmuller A, Schultheiss HP, Lamers J, Poller W (1999) Expression of coxsackie adenovirus receptor and alphav-integrin does not correlate with adenovector targeting in vivo indicating anatomical vector barriers. Gene Ther 6:1520–1535
Yamazaki T, Shiraishi S, Kishimoto K, Miura S, Ezaki O (2011) An increase in liver PPARgamma2 is an initial event to induce fatty liver in response to a diet high in butter: PPARgamma2 knockdown improves fatty liver induced by high-saturated fat. J Nutr Biochem 22(6):543–53
Lochmuller H, Jani A, Huard J, Prescott S, Simoneau M, Massie B, Karpati G, Acsadi G (1994) Emergence of early region 1-containing replication-competent adenovirus in stocks of replication-defective adenovirus recombinants (delta E1 + delta E3) during multiple passages in 293 cells. Hum Gene Ther 5:1485–1491
Kanegae Y, Makimura M, Saito I (1994) A simple and efficient method for purification of infectious recombinant adenovirus. Jpn J Med Sci Biol 47:157–166
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Yamazaki, T., Ezaki, O. (2013). Producing PPARgamma2 Knockdown in Mouse Liver. In: Badr, M., Youssef, J. (eds) Peroxisome Proliferator-Activated Receptors (PPARs). Methods in Molecular Biology, vol 952. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-155-4_7
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DOI: https://doi.org/10.1007/978-1-62703-155-4_7
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-154-7
Online ISBN: 978-1-62703-155-4
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