Viral vectors

Abstract

Constructs for delivery of sequences of interest to cells include a herpes virus latency active promoter (LAP) of the latency associated transcript (LAT) region. An internal ribosome entry site (IRES) is located downstream of the LAP, with a nucleotide sequence of interest downstream of the IRES. Stable, long-term expression, including export of mRNA to the cytoplasm and translation of the encoded polypeptide, is found in neuronal and non-neuronal cells.

1 . A nucleic acid construct including (i) a portion of the latency associated transcript (LAT) region of a herpes virus genome, which portion includes a latency active promoter (LAP), (ii) an internal ribosome entry site (IRES) downstream of the promoter, and (iii) a nucleotide sequence downstream of the IRES and heterologous to the LAT region.  
     
     
         2 . A nucleic acid construct according to    claim 1    wherein the heterologous nucleotide sequence encodes a polypeptide.  
     
     
         3 . A nucleic acid construct according to    claim 1    or    claim 2    which is part of a vector.  
     
     
         4 . A nucleic acid construct according to    claim 3    wherein the vector is a viral vector.  
     
     
         5 . A nucleic acid construct according to    claim 4    wherein the vector is a herpes viral vector.  
     
     
         6 . A nucleic acid construct according to    claim 5    wherein the LAT region is native to the herpes virus of the vector.  
     
     
         7 . A nucleic acid construct according to any of    claims 4    to    6    wherein the viral vector is replication defective.  
     
     
         8 . A nucleic acid construct according to any preceding claim wherein the herpes virus is herpes simplex virus  1  (HSV 1 ).  
     
     
         9 . A nucleic acid construct according to any preceding claim wherein the IRES is a picornavirus IRES.  
     
     
         10 . A cell containing a nucleic acid construct according to any preceding claim.  
     
     
         11 . A cell according to    claim 10    wherein the heterologous nucleotide sequence is being expressed.  
     
     
         12 . A cell according to    claim 10    or    claim 11    which is a nerve cell.  
     
     
         13 . A cell according to any of    claims 10    to    12    which is part of a mammal.  
     
     
         14 . A mammal having a cell according to any of    claims 10    to    12   .  
     
     
         15 . A mammal containing a nucleic acid construct according to any of    claims 1    to    9   .  
     
     
         16 . A method including introduction of a nucleic acid construct according to any of    claims 1    to    9    into a cell.  
     
     
         17 . A method according to    claim 16    wherein the cell is a nerve cell.  
     
     
         18 . A method according to    claim 16    or    claim 17    wherein said introduction takes place ex vivo.  
     
     
         19 . A method which includes causing or allowing expression of a heterologous nucleotide sequence in a nucleic acid construct according to any of the    claims 1    to    9    in a cell.  
     
     
         20 . A method according to    claim 19    wherein the cell is part of a mammal.  
     
     
         21 . A herpes virus including in its genome an internal ribosome entry site (IRES) downstream of the herpes virus latency active promoter (LAP) and a non-herpes virus nucleotide sequence downstream of the IRES.  
     
     
         22 . A herpes virus according to    claim 21    which is replication defective or attenuated.  
     
     
         23 . A cell containing a herpes virus according to    claim 21    or    claim 22   .  
     
     
         24 . A mammal having a cell according to    claim 23   .  
     
     
         25 . A method which includes administering to a mammal a nucleic acid construct according to any of    claims 1    to    9   .  
     
     
         26 . A method according to    claim 25    wherein a virus containing the nucleic acid construct is administered to the mammal.  
     
     
         27 . A method which includes administering to a mammal a cell according to any of    claims 10    to    12   .  
     
     
         28 . A method which includes administering to a mammal a herpes virus according to    claim 21    or    claim 22   .