title: Establishment of a novel triple-transgenic mouse: conditionally and liver-specifically expressing hepatitis C virus NS3/4A protease.
abstract: It is well known that NS3/4A protein plays crucial roles in the hepatitis C virus (HCV) replication. NS3/4A protein also results to virus-mediated immune evasion and persistence of infection through the interaction with host proteins. However, the lack of a suitable animal model hampers studies of HCV NS3/4A protein interaction with host proteins, which impacts immunopathology due to infection. Here, transgenic vector containing transcriptional regulation and Fluc reporter gene was constructed to conditionally express NS3/4A protein under the dual control of Tet-On regulatory system and Cre/LoxP gene-knockout system. NS3/4A transgenic founder mice were continuously crossed with Lap transgenic mice expressing reverse tetracycline-controlled transcriptional activator (rtTA), the NS3/4A/Lap double transgenic mouse lines with liver-specifically and conditionally expressing reporter (luciferase Fluc) under control of Tet-On system were established. The NS3/4A/Lap double transgenic mouse are mated with Lap/LC-1 double transgenic mouse with liver-specifically and conditionally expressing Cre recombinase under control of Tet-On system, NS3/4A/Lap/LC-1 triple transgenic mouse were generated. In vivo bioluminescent imaging, western blotting and immunohistochemical staining (IHS) was used to confirm that NS3/4A protein was strictly expressed in the liver of Doxycycline-induced triple transgenic mice. The results show that we established a triple-transgenic mouse model conditionally expressing the HCV NS3/4A protein under strict control of the Tet-On regulatory system and Cre/loxP system. This novel transgenic mouse model expressing NS3/4A in a temporally and spatially-specific manner will be useful for studying interactions between HCV NS3/4A protein and the host, also for evaluating NS3/4A protease inhibitors.
authors.affiliation: Department of Medical Microbiology and Parasitology, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: H Y
authors.initials: HY
authors.lastname: Lan
authors.forename: Y
authors.initials: Y
authors.lastname: Zhao
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.forename: M N
authors.initials: MN
authors.lastname: Sun
authors.forename: Y F
authors.initials: YF
authors.lastname: Lei
authors.forename: M
authors.initials: M
authors.lastname: Yao
authors.forename: X J
authors.initials: XJ
authors.lastname: Huang
authors.forename: J M
authors.initials: JM
authors.lastname: Zhang
authors.forename: Z K
authors.initials: ZK
authors.lastname: Xu
authors.forename: X
authors.initials: X
authors.lastname: Lü
authors.forename: W
authors.initials: W
authors.lastname: Yin
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers / Q000235:genetics; D004195:Disease Models, Animal; D055786:Gene Knockout Techniques; D005822:Genetic Vectors; D016174:Hepacivirus / Q000201:enzymology*; D007150:Immunohistochemistry; D019426:Integrases; D047908:Intracellular Signaling Peptides and Proteins; D008099:Liver / Q000378:metabolism*; D008156:Luciferases; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics* / Q000378:metabolism* / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D019426:Integrases; D047908:Intracellular Signaling Peptides and Proteins; D008156:Luciferases; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C455041:NS4A cofactor peptide, Hepatitis C virus; C045073:Cre recombinase