Highly permissive cell lines for hepatitis C virus RNA replication
This invention relates generally to cells and cell lines that are permissive for hepatitis C virus (HCV) replication, and methods and materials for making and using them. The subject cell line referred to as Huh-7.5 has the A.T.C.C. designation number PTA-8561, having been deposited on Aug. 1, 2007.
1. A method for producing a cell line permissive for HCV replication, the method comprising:
(a) providing an Huh-7.5 cell line on deposit with the ATCC as accession number PTA-8561 that further comprises a genomic or subgenomic HCV RNA capable of replication;
(b) obtaining a culture of said cell line that supports replication of said genomic or subgenomic HCV RNA;
(c) curing said cell line of (b) of HCV RNA, wherein said curing comprises treatment with an antiviral agent; and
(d) identifying sublines of said cured cell line of (b) that are permissive for HCV replication.
2. The method of claim 1 , wherein said agent is an antiviral cytokine.
3. The method of claim 2 , wherein said antiviral cytokine is interferon.
4. The method of claim 1 , wherein said cell line of (a) comprises a replicating subgenomic HCV RNA containing no adaptive mutations.
5. The method of claim 1 , wherein said cell line of (a) comprises a replicating subgenomic HCV RNA that comprises an adaptive mutation.
6. An Huh-7.5 cell line on deposit with the ATCC as accession number PTA-8561.
7. The cell line of claim 6 , wherein said cell line contains subgenomic HCV RNA.
8. The cell line of claim 7 , wherein said subgenomic HCV RNA comprises an adaptive mutation.
9. The cell line of claim 8 , wherein said adaptive mutation is S22041, S2204V, or S2204A, said position being identified by alignment with the genotype 1 b Con1 full-length HCV genome (SEQ ID NO:27) commencing with the core-coding region.
10. The cell line of claim 6 , wherein said cell line contains genomic HCV RNA.
11. The cell line of claim 10 , wherein said genomic HCV RNA comprises an adaptive mutation.
12. The cell line of claim 11 , wherein said adaptive mutation is S2204I, S2204V, or S2204A, said position being identified by alignment with the genotype 1b Con1 full-length HCV genome (SEQ ID NO:27) commencing with the core-coding region.