Control of gene expression

Abstract

A method of suppressing the expression of a selected gene in a eukaryotic cell the method comprising introducing into the cell (a) a polypeptide comprising a DNA binding portion which binds to a site at or associated with the selected gene which site is present in a plant or animal genome and a chromatin inactivation portion, or (b) a polynucleotide encoding said polypeptide.

1 . A method of suppressing the expression of a selected gene in a eukaryotic cell the method comprising introducing into the cell (a) a polypeptide comprising a nucleic acid binding portion which binds to a site at or associated with the selected gene which site is present in a eukaryotic genome and a chromatin inactivation portion, or (b) a polynucleotide encoding said polypeptide wherein the chromatin inactivation portion is selected from all or a N-CoR- or SMRT-binding part of PLZF or wherein the nucleic acid binding portion is selected from all or a DNA binding part of a nuclear receptor DNA binding protein.  
   
   
       2 . A method according to  claim 1  wherein when the nucleic acid binding portion is selected from a DNA binding portion of RARα the chromatin inactivation portion being other than a portion of PLZF protein and is other than a portion of PML protein; or wherein the nucleic acid binding portion is other than a DNA binding portion of the  Saccharomyces cerevisciae  GAL4 protein.  
   
   
       3 . A method according to  claim 1  wherein the nucleic acid binding portion is a DNA binding portion.  
   
   
       4 . A method according to  claim 1  wherein the nucleic acid binding portion is an RNA binding portion and the site present in a eukaryotic genome is a nascent RNA being transcribed from DNA.  
   
   
       5 . A method according to  claim 1  wherein when the nucleic acid binding portion is selected from all or a DNA binding part of a nuclear receptor DNA binding protein the chromatin inactivation portion facilitates histone deacetylation.  
   
   
       6 . A method according to  claim 3  wherein when the nucleic acid binding portion is selected from all or a DNA binding part of a nuclear receptor DNA binding protein the chromatin inactivation portion facilitates histone deacetylation.  
   
   
       7 . A method according to  claim 1  wherein when the nucleic acid binding portion is selected from all or a DNA binding part of a nuclear receptor DNA binding protein the chromatin inactivation portion is selected from all or a portion of a component of a histone deacetylation (HDAC) complex or all or a portion of a polypeptide which binds to or facilitates the recruitment of a HDAC complex.  
   
   
       8 . A method according to  claim 7  wherein the component of the HDAC complex or the polypeptide which binds to or facilitates the recruitment of a HDAC complex is selected from PLZF, N-CoR, SMRT, Sin3, SAP18, SAP30 and HDAC.  
   
   
       9 . A method according to  claim 8  wherein the chromatin inactivation portion is selected from all or a N-CoR- or SMRT-binding part of PLZF.  
   
   
       10 . A method according to  claim 8  wherein the chromatin inactivation portion is selected from all or an enzymatically active part of a HDAC.  
   
   
       11 . A method according to  claim 3  wherein when the chromatin inactivation portion is selected from all or a N-CoR- or SMRT-binding part of PLZF the DNA binding portion is selected from all or a DNA-binding part of a zinc-finger DNA binding protein or all or a DNA-binding part of a helix-turn-helix DNA binding protein.  
   
   
       12 . A method according to  claim 11  wherein the DNA binding portion is selected from all or a DNA-binding part selected from the group consisting of an animal or plant DNA binding protein and a bacterial or yeast DNA binding protein engineered to bind plant or animal genome.  
   
   
       13 . A method according to  claim 1  wherein the DNA binding portion is selected from all or a DNA binding part of a steroid hormone receptor protein.  
   
   
       14 . A method according to  claim 13  wherein the steroid hormone receptor protein is selected from all or a DNA-binding portion of estrogen receptor (ER) or all or a DNA-binding portion of androgen receptor (AR).  
   
   
       15 . A method according to  claim 4  wherein the RNA binding protein binds to nascent RNA expressed from proviral DNA.  
   
   
       16 . A method according to  claim 15  wherein the RNA binding protein is selected from tat or a tat-like protein or an RNA-binding portion thereof.  
   
   
       17 . A method according to  claim 1  wherein the nucleic acid binding portion and the chromatin inactivation portion are fused.  
   
   
       18 . A method according to  claim 1  wherein the eukaryotic cell is selected from an animal cell and is contained within an animal or a plant cell and is contained within a plant.  
   
   
       19 . A method according to  claim 1  wherein the expression of one or more selected genes in a human is suppressed.  
   
   
       20 . A method of treating a patient in need of suppression of the expression of a selected gene, the method comprising a step selected from 
 (a) administering to the patient an effective amount of a polypeptide comprising a nucleic acid binding portion which binds to a site at or associated with the selected gene and a chromatin inactivation portion; and    (b) administering to the patient an effective amount of a polynucleotide encoding a polypeptide comprising a nucleic acid binding portion which binds to a site at or associated with the selected gene and a chromatin inactivation portion.    
   
   
       21 . A composition selected from pharmaceutical compositions and compositions used in medicine selected from (a) a polypeptide comprising a nucleic acid binding portion which binds to a site at or associated with the selected gene which site is present in a eukaryotic genome and a chromatin inactivation portion, or (b) a polynucleotide encoding a polypeptide wherein the chromatin inactivation portion is selected from all or a N-CoR- or SMRT-binding part of PLZF or wherein the nucleic acid binding portion is selected from all or a DNA binding part of a nuclear receptor DNA binding protein.  
   
   
       22 . A composition according to  claim 21  including a pharmaceutically acceptable carrier.  
   
   
       23 . A composition according to  claim 21  wherein the composition is a polypeptide.  
   
   
       24 . A composition according to  claim 21  wherein the composition is a polynucleotide encoding a polypeptide.  
   
   
       25 . A polynucleotide encoding a polypeptide according to  claim 24  wherein the polynucleotide comprises a promoter operably linked to allow expression of the polypeptide.  
   
   
       26 . A polynucleotide according to  claim 25  wherein the promoter is an inducible promoter.  
   
   
       27 . A polynucleotide according to  claim 24  wherein said polynucleotide is included in a vector.  
   
   
       28 . A polynucleotide according to  claim 27  wherein said vector is selected from the group consisting of animal cell vectors and plant cell vectors.  
   
   
       29 . A polynucleotide according to  claim 27  wherein said vector is selected from the group consisting of viral vectors and plasmid vectors.  
   
   
       30 . A host cell comprising a polynucleotide according to  claim 21 .  
   
   
       31 . A host cell according to  claim 30  wherein said host cell is selected from the group consisting of bacterial cells, animal cells and plant cells.