Znf206: a novel regulator of embryonic stem cell self-renewal and pluripotency

Abstract

We have identified ZNF206, a novel repressor of human embryonic stem cell (hESC) differentiation. Repressing extra-embryonic endoderm development preserves the pluripotent state of human embryonic stem cells, and, conversely down-regulating expression of ZNF206 in hESCs causes them to upregulate the expression of genes associated with the extra-embryonic endodermal lineage, down-regulate genes associated with the pluripotent state, and may lead to the further emergence of genes associated with even more differentiated lineages and phenotypes.

1 . An isolated polynucleotide comprising a sequence that has at least 90% nucleic acid sequence identity to a native ZNF206 polynucleotide and that hybridizes selectively to the native ZNF206 polynucleotide. 
     
     
         2 - 3 . (canceled) 
     
     
         4 . The polynucleotide of  claim 1  comprising a sequence at least 100 nucleotides in length that has at least 90% nucleic acid sequence identity to a native ZNF206 polynucleotide. 
     
     
         5 - 6 . (canceled) 
     
     
         7 . The polynucleotide of  claim 1  comprising at least 15 contiguous nucleotides of a native ZNF206 polynucleotide, wherein the isolated polynucleotide hybridizes selectively to a native ZNF206 polynucleotide. 
     
     
         8 - 9 . (canceled) 
     
     
         10 . The isolated polynucleotide of  claim 7  comprising a full-length protein-coding sequence of a native ZNF206 mRNA or cDNA. 
     
     
         11 . The isolated polynucleotide of  claim 1  that encodes a polypeptide that has ZNF206 activity. 
     
     
         12 - 19 . (canceled) 
     
     
         20 . An isolated polypeptide of at least 11 amino acids that comprises at least 4 contiguous amino acids of a native ZNF206 polypeptide, and, that when introduced into a mammal, elicits production of an antibody that binds selectively to a native ZNF206 polypeptide. 
     
     
         21 - 36 . (canceled) 
     
     
         37 . The isolated polypeptide of  claim 20  that has ZNF206 activity. 
     
     
         38 . (canceled) 
     
     
         39 . An antibody that binds selectively to a native ZNF206 polypeptide. 
     
     
         40 . The antibody of  claim 39 , wherein the antibody is selected from the group consisting of monoclonal antibody, polyclonal antibody, chimeric antibody, humanized antibody, and single chain antibody. 
     
     
         41 - 48 . (canceled) 
     
     
         49 . A method of detecting the presence of a ZNF206 polynucleotide in a sample comprising the ZNF206 polynucleotide, the method comprising contacting the sample with (a) a probe or primer comprising a polynucleotide sequence that binds selectively to the ZNF206 polynucleotide and detecting binding of the probe or primer to the ZNF206 mRNA, or (b) contacting the sample with an antibody that binds selectively to the ZNF206 polypeptide and detecting binding of the antibody to the ZNF206 polypeptide. 
     
     
         50 . The method of  claim 49  comprising (a) contacting the sample with a first primer that comprises the polynucleotide sequence that hybridizes selectively to the ZNF206 mRNA and a second primer comprising a polynucleotide sequence that hybridizes to the ZNF206 mRNA, (b) performing an amplification reaction to produce an amplification product that indicates the presence of the ZNF206 mRNA in the sample, and (c) detecting the amplification product. 
     
     
         51 - 56 . (canceled) 
     
     
         56 . A method of assessing the pluripotency of a cell comprising (a) measuring ZNF206 polypeptide or polynucleotide levels in a sample comprising the cell, and (b) comparing the ZNF206 polypeptide or polynucleotide levels in the sample to a reference. 
     
     
         57 . The method of  claim 56  comprising measuring the ZNF206 polypeptide level in the cell by contacting the sample with an antibody that binds selectively to ZNF206 polypeptide, and measuring binding of the antibody to ZNF206 polypeptide in the sample. 
     
     
         58 - 60 . (canceled) 
     
     
         61 . The method of  claim 56  comprising measuring the ZNF206 mRNA level in the cell by contacting the sample with a probe or primer that hybridizes selectively to ZNF206 mRNA and measuring hybridization of the probe or primer to the ZNF206 mRNA in the sample. 
     
     
         62 - 69 . (canceled) 
     
     
         70 . A method of promoting differentiation of a cell by reducing ZNF206 expression of the cell, the method comprising expressing in the cell a vector comprising (a) a promoter suitable for expression in the cell operably linked to (b) an isolated polynucleotide comprising a sequence that has at least 90% nucleic acid sequence identity to a native ZNF206 polynucleotide and that hybridizes selectively to the native ZNF206 polypeptide, wherein expression of the polynucleotide in the cell causes a reduction in ZNF206 polypeptide levels in the cell. 
     
     
         71 . A method of diagnosing a cancer characterized by elevated levels of ZNF206 comprising (a) obtaining a sample comprising a cell, (b) determining ZNF206 polypeptide or polynucleotide levels in the sample, and (c) comparing the ZNF206 polypeptide or polynucleotide levels in the sample with a reference. 
     
     
         72 . A method of treating a cancer characterized by elevated levels of ZNF206 comprising administering to a patient in need of such treatment a composition comprising a vector comprising (a) a promoter suitable for expression in a cell of the patient operably linked to (b) an isolated polynucleotide comprising a sequence at least 100 nucleotides in length that has at least 90% nucleic acid sequence identity to a native ZNF206 polynucleotide, wherein expression of the polynucleotide in the cell reduces ZNF206 polypeptide levels in the cell. 
     
     
         73 - 79 . (canceled)