Controlled gene expression methods
Methods and compositions for controlling gene expression are disclosed.
1. A nucleic acid molecule encoding a polypeptide comprising a DNA binding domain and a looping factor, wherein said looping factor is a fragment of LIM domain binding 1 (Ldb1), wherein said fragment of Ldb1 comprises an amino acid sequence that has at least 90% identity with amino acids 1 to 200 of SEQ ID NO: 1.
2. The nucleic acid molecule of claim 1 , wherein said DNA binding domain and said looping factor are linked through a covalent bond or an amino acid linker.
3. The nucleic acid molecule of claim 1 , wherein said Ldb1 is human.
4. The nucleic acid molecule of claim 1 , wherein said fragment of Ldb1 comprises amino acids 1 to 200 of SEQ ID NO: 1.
5. The nucleic acid molecule of claim 1 , wherein said DNA binding domain comprises a zinc finger protein or a transcription activator-like effector protein.
6. The nucleic acid molecule of claim 1 , wherein said DNA binding domain specifically binds a target sequence in the promoter of a gene of interest.
7. The nucleic acid molecule of claim 6 , wherein said promoter is a globin promoter.
8. The polypeptide encoded the nucleic acid molecule of claim 1 .
9. A vector comprising the nucleic acid molecule of claim 1 .
10. A composition comprising at least one polypeptide of claim 8 and at least one pharmaceutically acceptable carrier.
11. A composition comprising at least one nucleic acid molecule of claim 1 and at least one pharmaceutically acceptable carrier.
12. A method of modulating the expression of a gene of interest in a cell, said method comprising expressing the nucleic acid molecule of claim 1 in the cell.
13. The method of claim 12 , wherein said method comprises expressing a first and second nucleic acid molecule in the cell,
wherein said first nucleic acid molecule encodes a first polypeptide comprising a first DNA binding domain and a first looping factor, wherein said first looping factor is a fragment of LIM domain binding 1 (Ldb1), and wherein said first DNA binding domain specifically binds to a target sequence in the promoter of said gene of interest, and
wherein said second nucleic acid molecule encodes a second polypeptide comprising a second DNA binding domain and a second looping factor, wherein said second looping factor is a fragment of LIM domain binding 1 (Ldb1), and wherein said second DNA binding domain specifically binds to a target sequence in the locus control region of said gene of interest.
14. The method of claim 13 , wherein said fragment of Ldb1 comprises amino acids 1 to 200 of SEQ ID NO: 1.
15. The nucleic acid molecule of claim 7 , wherein said promoter is the fetal gamma globin promoter.
16. The nucleic acid molecule of claim 1 , wherein said looping factor consists of amino acids 1 to 200 of SEQ ID NO: 1.
17. The nucleic acid molecule of claim 1 , wherein said fragment of Ldb1 comprises an amino acid sequence that has at least 97% identity with amino acids 1 to 200 of SEQ ID NO: 1.