Tau protein-binding dna aptamers with capture/detection and therapeutic utilities in tauopathy-related neurodegenerative disorders

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, and SEQ ID NO:
• 11;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 1.

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, and SEQ ID NO:
• 16;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 2.

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 17, SEQ ID NO:
• 18, and SEQ ID NO:
• 19;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 3.

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 20, SEQ ID NO:
• 21, and SEQ ID NO:
• 22;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 4.

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 23, SEQ ID NO:
• 24, and SEQ ID NO:
• 25;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 5.

A DNA aptamer comprising a nucleic acid sequence that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;

• wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NO:
• 26 and SEQ ID NO:
• 27;
• and wherein the nucleic acid sequence is a truncated fragment of a nucleic acid sequence set forth in SEQ ID NO:
• 6.

A composition comprising a DNA aptamer that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein, wherein the DNA aptamer comprises a nucleic acid sequence comprising 66 nucleotides and is selected from the group consisting of SEQ ID NO:

• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, and SEQ ID NO:
• 7.

A composition comprising a DNA aptamer that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein, wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:

• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.

A DNA aptamer conjugate comprising:

• a signaling moiety conjugated to a DNA aptamer that is capable of binding to a tau protein at a phosphorylatable site of the tau protein,
• wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.

A DNA aptamer conjugate comprising:

• a DNA aptamer that is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein, and an azido-containing cell penetrating peptide linked to the DNA aptamer,
• wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.

A method for cell-based imaging of phosphorylated tau, non-phosphorylated tau, and/or total tau in a cell comprising:

• introducing a DNA aptamer conjugate into a cell in a cell culture medium;
• and
• detecting the binding of the DNA aptamer conjugate to a tau protein in the cell;
• wherein the DNA aptamer conjugate comprises a modified DNA aptamer that is linked with an azido-containing cell penetrating peptide;
• wherein the modified DNA aptamer is capable of specifically binding to a tau protein at a phosphorylatable site of the tau protein;
• and
• wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.

A method for in vivo imaging of phosphorylated tau, non-phosphorylated tau, and/or total tau in a subject comprising:

• delivering a composition comprising a DNA aptamer conjugate into a subject in need thereof;
• and
• detecting the binding of the DNA aptamer conjugate to a tau protein in the subject;
• wherein the DNA aptamer conjugate comprises a modified DNA aptamer that is linked with an azido-containing cell penetrating peptide;
• and
• wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.

A method for in vivo imaging of phosphorylated tau, non-phosphorylated tau, and/or total tau in a subject comprising:

• delivering a composition comprising a DNA aptamer conjugate into a subject in need thereof;
• and
• detecting the binding of the DNA aptamer conjugate to a tau protein in the subject;
• wherein the DNA aptamer conjugate comprises a DNA aptamer conjugated to a moiety, wherein the moiety is a reporter molecule which can be a molecular beacon, fluorescent tag, a radioisotope for positron emission tomography (PET), single-photon emission computed tomography (SPECT), and/or contrast-agent-based MRI;
• and
• wherein the DNA aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8, SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, SEQ ID NO:
• 12, SEQ ID NO:
• 13, SEQ ID NO:
• 14, SEQ ID NO:
• 15, SEQ ID NO:
• 16, SEQ ID NO:
• 17, SEQ ID NO:
• 18, SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 21, SEQ ID NO:
• 22, SEQ ID NO:
• 23, SEQ ID NO:
• 24, SEQ ID NO:
• 25, SEQ ID NO:
• 26, and SEQ ID NO:
• 27.