Reagents for treatment of hepatitis b virus (hbv) infection and use thereof

Abstract

This disclosure relates to RNA interference (RNAi) reagents for treatment of hepatitis B virus (HBV) infection, compositions comprising same, and use thereof to treat individuals infected with HBV.

1 - 41 . (canceled) 
     
     
         42 . A DNA-directed RNA interference (ddRNAi) construct comprising:
 (a) a first nucleic acid comprising a DNA sequence which encodes a short hairpin RNA (shRNA) comprising an effector sequence of at least 19 nucleotides in length which is substantially complementary to a RNA transcript encoded by the sequence set forth in SEQ ID NO: 9; and   (b) a second nucleic acid comprising a DNA sequence which encodes a shRNA comprising an effector sequence of at least 19 nucleotides in length which is substantially complementary to a RNA transcript encoded by the sequence set forth in SEQ ID NO:   
     
     
         58 . 
     
     
         43 . The ddRNAi construct of  claim 42 , wherein:
 (a) the first nucleic acid comprises a DNA sequence which encodes a shRNA comprising an effector sequence which is substantially complementary to a RNA transcript encoded by the sequence set forth in SEQ ID NO: 48; and   (b) the second nucleic acid comprises a DNA sequence which encodes a shRNA comprising an effector sequence which is substantially complementary to a RNA transcript encoded by the sequence set forth in SEQ ID NO: 58.   
     
     
         44 . The ddRNAi construct of  claim 43 , wherein:
 (a) the first nucleic acid comprises a DNA sequence encoding a shRNA comprising an effector sequence set forth in SEQ ID NO:47 and an effector complement sequence set forth in SEQ ID NO:48; and   (b) the second nucleic acid comprises a DNA sequence encoding a shRNA comprising an effector sequence set forth in SEQ ID NO:57 and an effector complement sequence set forth in SEQ ID NO:58.   
     
     
         45 . The ddRNAi construct according to  claim 44 , wherein:
 (a) the first nucleic acid comprises a DNA sequence encoding a shRNA comprising or consisting of the sequence set forth in SEQ ID NO:92; and   (b) the second nucleic acid comprises a DNA sequence encoding a shRNA comprising or consisting of the sequence set forth in SEQ ID NO:101.   
     
     
         46 . The ddRNAi construct according to  claim 42 , comprising a RNA pol III promoter upstream of each nucleic acid encoding a shRNA. 
     
     
         47 . The ddRNAi construct according to  claim 46 , wherein each RNA pol III promoter is selected from a U6 and a H1 promoter. 
     
     
         48 . The ddRNAi construct according to  claim 47 , wherein the U6 promoter is a U6-1 promoter, U6-8 promoter or U6-9 promoter. 
     
     
         49 . An expression vector comprising the ddRNAi construct of  claim 42 . 
     
     
         50 . The expression vector of  claim 49 , wherein:
 the expression vector is a plasmid or minicircle; or   (ii) the expression vector is a viral vector selected from the group consisting of: an adeno-associated viral (AAV) vector, a retroviral vector, an adenoviral (AdV) vector and a lentiviral (LV) vector;
 optionally wherein the or each expression vector is complexed with a cationic DNA binding polymer. 
   
     
     
         51 . The expression vector of  claim 49 , wherein the expression vector is an adeno-associated viral (AAV) vector. 
     
     
         52 . A composition comprising a ddRNAi construct according to  claim 42  and one or more pharmaceutically acceptable carriers, optionally wherein the ddRNAi construct is comprised within an expression vector. 
     
     
         53 . The composition of  claim 52 , wherein the expression vector is an adeno-associated viral (AAV) vector. 
     
     
         54 . A method of treating Hepatitis B virus (HBV) infection in a subject, said method comprising administering to the subject a composition of  claim 52 . 
     
     
         55 . The method according to  claim 54 , wherein the subject is suffering from chronic HBV infection. 
     
     
         56 . The method of  claim 54 , wherein administering the composition to the subject reduces HBV viral load in the subject and/or reduces severity of symptoms associated with HBV infection in the subject and/or reduces infectivity of HBV in the subject.