Detectable nucleic acid tag
View Patent ↗Provided herein are nucleic acid tags that are linked to, or capable of linking to, a protein of interest. In particular, the nucleic acid tags are oligonucleotides comprising a reporter function and a protein tagging function. Also provided herein, are nucleic acid tag compositions, kits and methods of use thereof.
1. A composition comprising a nucleic acid oligomer bound to a fusion protein through a nucleic acid-interacting motif in said fusion protein, wherein the fusion protein comprises:
(a) a first protein domain comprising a human protein of interest, and (b) a second, different protein domain comprising said nucleic acid-interacting motif,
wherein the nucleic acid oligomer is between about 50 and about 500 nucleotides in length and
comprises:
(c) a first nucleic acid sequence that is a PCR amplicon sequence not having a recognition sequence for said nucleic acid-interacting motif of said fusion protein, and
(d) a second nucleic acid sequence having a recognition sequence specifically bound to said nucleic acid-interacting motif of said fusion protein, wherein the first nucleic acid sequence is heterologous to the second nucleic acid sequence, said nucleic acid oligomer further comprising the nucleotide sequence depicted in SEQ ID NO:1; SEQ ID NO:2; SEQ ID NO:3 or SEQ ID NO:4.
2. The composition of claim 1 , wherein the nucleic acid oligomer is a double-stranded DNA.
3. The composition of claim 1 , wherein the nucleic acid-interacting motif is a DNA-binding domain.
4. A composition comprising a nucleic acid oligomer bound to a fusion protein through a nucleic acid-interacting motif therein, wherein the fusion protein comprises:
(a) a first protein domain comprising a human protein of interest, and
(b) a second, different protein domain comprising said nucleic acid-interacting motif;
wherein the nucleic acid oligomer is between about 50 and about 500 nucleotides in length and
comprises:
(c) a first nucleic acid sequence that is a PCR amplicon sequence not having a recognition sequence for said nucleic acid-interacting motif of said fusion protein, and
(d) a second nucleic acid sequence having a recognition sequence for specifically binding to said nucleic acid-interacting motif of said fusion protein, wherein the oligomer is radiolabeled, fluorescently labeled or biotinylated, said nucleic acid oligomer further comprising the nucleotide sequence depicted in SEQ ID NO:1; SEQ ID NO:2; SEQ ID NO:3 or SEQ ID NO:4.
5. The composition of claim 1 or 4 , wherein the protein of interest is a transmembrane protein, transmembrane ion channel protein, ligand gated ion channel protein, nuclear hormone receptor protein, extracellular signaling molecule or factor, cytokine, growth factor, hormone, enzyme, antibody or small chain variable fragment (scFv).
6. The composition of claim 1 or 4 , wherein the protein of interest is a kinase in the activated or inactivated state.
7. The composition of claim 6 , wherein the kinase is a human kinase.
8. The composition of claim 6 , wherein the kinase is a non-receptor tyrosine kinase.
9. The composition of claim 8 , wherein the non-receptor tyrosine kinase is a member of the ABL, ACK, CSK, MATK, FAK, PYK2, FES, FRK, JAK, SRC-A, SRC-B, TEC, or SYK family of tyrosine kinases.
10. The composition of claim 6 , wherein the kinase is a receptor tyrosine kinase.
11. The composition of claim 10 , wherein the receptor tyrosine kinase is member of the ALK, AXL, DDR, EGFR, EPH, FGFR, INSR, MET, MUSK, PDGFR, PTK7, RET, ROR, ROS, RYK, TIE, TRK, VEGFR, AATYK, or SuRTK106 family of tyrosine kinases.
12. A nucleic acid oligomer comprising the nucleotide sequence depicted in SEQ ID NO:1; SEQ ID NO:2; SEQ ID NO:3 or SEQ ID NO:4.
13. The composition of claim 1 , wherein the second domain comprises the amino acid sequence depicted in SEQ ID NO: 5 or SEQ ID NO: 6.
14. The composition of claim 1 wherein said protein of interest is a transmembrane protein.
15. The composition of claim 1 wherein said recognition sequence of said nucleic acid oligomer specifically binds to a transcription factor.
16. The composition of claim 15 wherein said recognition sequence specifically binds to a transcription factor which is NFκB or GAL4.
17. The composition of claim 1 further comprising an immobilized ligand bound to said fusion protein.
18. The composition of claim 4 further comprising an immobilized ligand bound to said fusion protein.