Modulation of transthyretin expression

Abstract

Compounds, compositions and methods are provided for modulating the expression of transthyretin. The compositions comprise oligonucleotides, targeted to nucleic acid encoding transthyretin. Methods of using these compounds for modulation of transthyretin expression and for diagnosis and treatment of diseases and conditions associated with expression of transthyretin are provided.

1 .- 18 . (canceled) 
     
     
         19 . A detection method comprising administering to a subject a modulator of human transthyretin expression, detecting nucleic acid or protein levels of transthyretin in a biological sample from said subject, wherein the biological sample is serum, adipose tissue, liver, body fluid, or a tissue or organ of an animal. 
     
     
         20 . The detection method of  claim 19 , wherein the nucleic acid of transthyretin is detected by polymerase chain reaction (PCR). 
     
     
         21 . The detection method of  claim 20 , wherein detecting the nucleic acid of transthyretin by PCR comprises contacting a preferred target segment of a nucleic acid molecule encoding human transthyretin with an oligonucleotide probe, a forward primer, and a reverse primer, wherein the probe comprises two fluorescent dyes and the forward primer and reverse primer flank the preferred target segment of the nucleic acid molecule encoding human transthyretin. 
     
     
         22 . The detection method of  claim 21 , wherein the forward primer comprises SEQ ID NO: 5. 
     
     
         23 . The detection method of  claim 21 , wherein the reverse primer comprises SEQ ID NO: 6. 
     
     
         24 . The detection method of  claim 21 , wherein the probe comprises SEQ ID NO: 
     
     
         25 . The detection method of  claim 24 , wherein the two fluorescent dyes are FAM and TAMRA. 
     
     
         26 . The detection method of  claim 19 , wherein the nucleic acid or protein level of transthyretin is detected by Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA) or fluorescence-activated cell sorting (FACS). 
     
     
         27 . The method of  claim 19 , wherein the modulator of transthyretin expression comprises an antisense compound, an antisense oligonucleotide, a ribozyme, a DNA oligonucleotide, an RNA oligonucleotide, or an RNA oligonucleotide having at least a portion of said RNA oligonucleotide capable of hybridizing with RNA to form an oligonucleotide-RNA duplex. 
     
     
         28 . The method of  claim 20 , wherein the antisense compound is single-stranded. 
     
     
         29 . The method of  claim 21 , wherein the antisense compound comprises a modified oligonucleotide 20 to 25 nucleobases in length having at least one modified sugar moiety. 
     
     
         30 . The method of  claim 22 , wherein the modified oligonucleotide is at least 95% complementary to SEQ ID NO: 4. 
     
     
         31 . A diagnostic method for identifying a transthyretin disease state in a subject comprising identifying the presence of human transthyretin in the subject by PCR comprising contacting a preferred target segment of a nucleic acid molecule encoding human transthyretin with an oligonucleotide probe, a forward primer, and a reverse primer, wherein the probe comprises two fluorescent dyes and the forward primer and reverse primer flank the preferred target segment of the nucleic acid molecule encoding human transthyretin. 
     
     
         32 . The diagnostic method of  claim 31 , wherein the forward primer comprises SEQ ID NO: 5. 
     
     
         33 . The diagnostic method of  claim 31 , wherein the reverse primer comprises SEQ ID NO: 6. 
     
     
         34 . The diagnostic method of  claim 31 , wherein the probe comprises SEQ ID NO: 7. 
     
     
         35 . The diagnostic method of  claim 34 , wherein the two fluorescent dyes are FAM and TAMRA. 
     
     
         36 . The diagnostic method of  claim 31 , wherein the forward primer comprises SEQ ID NO: 5, the reverse primer comprises SEQ ID NO: 6, and the probe comprises SEQ ID NO: 7. 
     
     
         37 . The diagnostic method of  claim 36 , wherein the two fluorescent dyes are FAM and TAMRA. 
     
     
         38 . The diagnostic method of  claim 31 , wherein the preferred target segment is present in a biological sample from the subject, wherein the biological sample is serum, adipose tissue, liver, body fluid, or a tissue or organ of the subject.