Inhibition of gene expression using polynucleotide analogues

Abstract

The invention provides sequence specific polynucleotide analogues and methods for determining the function of a nucleic acid of known sequence.

1 - 31 . (canceled)  
     
     
         32 . A method for determining a phenotype associated with a selected nucleic acid in a teleost embryo or egg giving rise to said embryo, wherein said embryo or egg is of a teleost species that undergoes meroblastic cleavage, said method comprising: 
 (a) contacting said teleost embryo or egg giving rise to said embryo with a morpholino-modified polynucleotide analogue that targets said selected nucleic acid; and    (b) detecting an altered phenotype in said teleost embryo or egg, or embryo developing from said egg, wherein said altered phenotype is associated with reduced expression or altered function of said selected nucleic acid.    
     
     
         33 . The method of  claim 32 , wherein said selected nucleic acid is a maternal or zygotic nucleic acid.  
     
     
         34 . The method of  claim 32 , wherein said altered phenotype is observed from fertilization, through organogenesis, to the completion of embryogenesis.  
     
     
         35 . The method of  claim 32 , said method further comprising contacting said embryo or egg giving rise to said embryo with a rescue mRNA, wherein said rescue mRNA encodes a polypeptide whose expression is reduced by said analogue, and wherein said rescue mRNA is present in an amount sufficient for expression of said polypeptide at a level comparable to that of a teleost embryo, or egg giving rise to said embryo, that is free of said analogue.  
     
     
         36 . A method for determining if a phenotype mediated by a polynucleotide analogue in a teleost organism is sequence-specific, said method comprising: 
 a) contacting a first teleost embryo or teleost egg with said polynucleotide analogue;    b) assessing the phenotype of said first teleost embryo or egg, or a teleost embryo developing from said egg, subsequent to step (a);    c) contacting a second teleost embryo or teleost egg with (i) said polynucleotide analogue and (ii) a rescue mRNA molecule;    d) assessing the phenotype of said second teleost embryo or egg, or a teleost embryo developing from said egg, subsequent to step (c); and    e) comparing the results of (b) and (d), wherein a phenotype detected in (b) that is not detected in (d) indicates that said phenotype detected in (b) is sequence-specific.    
     
     
         37 - 67 . (canceled)