tailieunhanh - Báo cáo khoa học: Hairpin ribozymes in combination with siRNAs against highly conserved hepatitis C virus sequence inhibit RNA replication and protein translation from hepatitis C virus subgenomic replicons

Chronic hepatitis C virus (HCV) infection is a clinically important liver disease with limited therapeutic options in a significant proportion of patients. Therefore, novel efficient therapeutic agents are needed. Because the 5¢- and 3¢-untranslated regions (UTRs) of HCV are highly conserved and functionally important for HCV replication, they are attractive targets for RNA-cleaving ribozymes or small interfering RNAs (siRNAs). | iFEBS Journal Hairpin ribozymes in combination with siRNAs against highly conserved hepatitis C virus sequence inhibit RNA replication and protein translation from hepatitis C virus subgenomic replicons Dominik Jarczak1 z Mortimer Korf1 z Carmela Beger2 Michael P. Manns1 and Martin Kruger1 1 Department of Gastroenterology Hepatology and Endocrinology Medizinische Hochschule Hannover Germany 2 Institute of Celland Molecular Pathology Medizinische Hochschule Hannover Germany Keywords constitutive transport element hairpin ribozyme hepatitis C virus small interfering RNA subgenomic replicon Correspondence M. Kruger Department of Gastroenterology Hepatology and Endocrinology Medizinische Hochschule Hannover Carl-Neuberg-Str. 1 D-30625 Hannover Germany Fax 01149 511532 4896 Tel 01149 511532 3305 E-mail Both authors contributed equally to this work Received 17 May 2005 revised 5 September 2005 accepted 22 September 2005 doi Chronic hepatitis C virus HCV infection is a clinically important liver disease with limited therapeutic options in a significant proportion of patients. Therefore novel efficient therapeutic agents are needed. Because the 5 - and 3 -untranslated regions UTRs of HCV are highly conserved and functionally important for HCV replication they are attractive targets for RNA-cleaving ribozymes or small interfering RNAs siRNAs . In this study hairpin ribozymes Rz targeting HCV 5 - and 3 -UTR sequences were expressed from a retroviral vector transcript under control of two different RNA polIII promoters tRNAVal U6 . Ribozymes were evaluated in monocistronic subgenomic I389 hyg-ub NS3-3 HCV replicon cells as single agents or in combination with siRNAs against HCV 5 - or 3 -UTR recently demonstrated to inhibit HCV replicons. Additionally ribozyme constructs were generated with the 3 -terminus of the ribozyme flanked by constitutive transport element CTE sequences an RNA motif that has previously

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