Method of detection and treatment of colon cancer
View Patent ↗The present invention is based, in part, on the discovery that colon carcinoma, carcinogenesis, or the predisposition thereto is associated with the level of Wnt2, Wnt5, BMP6, and Fz receptors and the full-length and dominant negative form of LEF1.
1. An isolated DNA molecule comprising a nucleic acid sequence encoding a dominant negative (LEF1 DN ) form of LEF-1 form encoded by SEQ ID NO:1, wherein the LEF1 DN lacks the N-terminal 115 amino acid residues of LEF-1, and represses the ability of full-length LEF-1 protein and a β-catenin to activate a reporter gene, and wherein the LEF1 DN consists of amino acids 116-399 of SEQ ID NO:2.
2. The isolated molecule of claim 1 , further comprising in operable linkage an LEF-1 promoter which is contained in a 232 nucleotide EspI-XhoI fragment of the nucleic acid sequence of in SEQ ID NO:3.
3. An isolated DNA molecule having regulatory activity and comprising a LEF-1 promoter which is contained in a 232 nucleotide EspI-XhoI fragment of the nucleic acid sequence of in SEQ ID NO:3 operably linked to a sequence heterologous to the nucleic acid encoding full length LEF-1.
4. The isolated DNA molecule of claim 3 , wherein the isolated DNA molecule comprises nucleotides 13-250 of SEQ ID NO:3.
5. The isolated DNA molecule of claim 3 , wherein the heterologous sequence encodes a marker.
6. The isolated DNA molecule of claim 5 , wherein the marker is green fluorescent protein or luciferase.
7. A vector comprising the isolated DNA molecule of claim 1 .
8. The vector of claim 7 , wherein the vector is an expression vector.
9. An isolated host cell comprising the vector of claim 7 .
10. An isolated host cell comprising the vector of claim 8 .
11. A method for producing LEF1 DN comprising:
a) culturing the host cell of claim 10 in culture medium;
b) recovering the LEF1 DN from the culture medium; and
c) purifying the LEF1 DN from the culture medium.
12. A vector comprising the isolated DNA molecule of claim 3 .
13. The vector of claim 12 , wherein the vector is an expression vector.
14. An isolated host cell comprising the vector of claim 12 .
15. An isolated host cell comprising the vector of claim 13 .
16. A method for producing a protein encoded by the heterologous sequence comprising:
a) culturing the host cell of claim 15 in culture medium;
b) recovering the protein from the culture medium; and
c) purifying the protein from the culture medium.