Establishment of infectious HCV virion-producing cells with newly designed full-genome replicon RNA.
Hepatitis C virus (HCV) replicon systems enable in-depth analysis of the life cycle of HCV. However, the previously reported full-genome replicon system is unable to produce authentic virions. On the basis of these results, we constructed newly designed full-genomic replicon RNA, which is composed of the intact 5'-terminal-half RNA extending to the NS2 region flanked by an extra selection marker gene. Huh-7 cells harboring this full-genomic RNA proliferated well under G418 selection and secreted virion-like particles into the supernatant. These particles, which were round and 50 nm in diameter when analyzed by electron microscopy, had a buoyant density of 1.08 g/mL that shifted to 1.19 g/mL after NP-40 treatment; these figures match the putative densities of intact virions and nucleocapsids without envelope. The particles also showed infectivity in a colony-forming assay. This system may offer another option for investigating the life cycle of HCV.
Pharmacology Research Laboratories I, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida-cho, Aoba-ku, Yokohama, Japan.
Masaaki
M
Arai
Hidenori
H
Suzuki
Yoshimi
Y
Tobita
Asako
A
Takagi
Koichi
K
Okamoto
Atsunori
A
Ohta
Masayuki
M
Sudoh
Michinori
M
Kohara
D002460:Cell Line; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D016366:Open Reading Frames; D012367:RNA, Viral / Q000235:genetics*; D012093:Replicon; D014771:Virion / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure; D014779:Virus Replication / Q000235:genetics
D012367:RNA, Viral
J Virol. 2006 May;80(9):4633-9
16611923
J Biol Chem. 2004 May 21;279(21):22371-6
14990575
Science. 1999 Jul 2;285(5424):110-3
10390360
J Virol. 2004 Aug;78(15):7904-15
15254163
Gastroenterology. 1999 Mar;116(3):636-42
10029622
J Virol. 2002 Dec;76(24):13001-14
12438626
J Med Virol. 2001 Jul;64(3):334-9
11424123
J Virol. 2002 Mar;76(6):2997-3006
11861865
J Virol. 1997 Jan;71(1):248-58
8985344
J Virol. 1995 Aug;69(8):4797-806
7541843
Gene Ther. 2006 Jun;13(11):883-92
16496015
Proc Natl Acad Sci U S A. 2006 May 9;103(19):7408-13
16651538
Gastroenterology. 2003 Dec;125(6):1808-17
14724833
Br J Haematol. 2003 Jun;121(6):904-12
12786802
Science. 2005 Jul 22;309(5734):623-6
15947137
Virus Res. 2006 Dec;122(1-2):1-10
16979254
Proc Natl Acad Sci U S A. 2006 Feb 14;103(7):2310-5
16461899
J Biol Chem. 2004 Apr 9;279(15):14531-41
14747471
J Virol. 2001 Sep;75(18):8516-23
11507197
J Virol. 1998 Oct;72(10):7972-7
9733835
Nat Med. 2005 Jul;11(7):791-6
15951748
Hepatol Res. 2007 Jun;37(6):433-43
17437527
J Virol. 1994 Jun;68(6):3925-33
8189529
Proc Natl Acad Sci U S A. 2005 Jun 28;102(26):9294-9
15939869
Virology. 2000 Aug 1;273(2):391-9
10915610
Science. 1998 Oct 30;282(5390):938-41
9794763
Biochem Biophys Res Commun. 2002 May 10;293(3):993-9
12051758
Hepatology. 1994 Feb;19(2):296-302
8294087
J Virol. 2002 Apr;76(8):4008-21
11907240
Science. 1989 Apr 21;244(4902):359-62
2523562