Controlled gene expression methods
Methods and compositions for controlling gene expression are disclosed.
1. A method of inhibiting a disease or disorder in a subject comprising administering to said subject a composition comprising i) 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), and ii) at least one pharmaceutically acceptable carrier,
wherein said fragment of Ldb1 comprises an amino acid sequence having at least 90% identity with amino acids 1 to 200 of SEQ ID NO: 1.
2. The method of claim 1 , wherein said disease or disorder is sickle cell anemia or beta-thalassemia and wherein said DNA binding domain specifically binds a target sequence in a globin promoter.
3. The method of claim 2 , wherein said DNA binding domain specifically binds a target sequence in the fetal (gamma) gene promoter.
4. The method of claim 1 , wherein said DNA binding domain and said looping factor are linked through a covalent bond or an amino acid linker.
5. The method of claim 1 , wherein said Ldb1 is human.
6. The method of claim 1 , wherein said fragment of Ldb1 comprises amino acids 1 to about 200 of SEQ ID NO: 1.
7. The method of claim 1 , wherein said DNA binding domain comprises a zinc finger protein or a transcription activator-like effector protein.
8. The method of claim 1 , wherein said DNA binding domain specifically binds a target sequence in the promoter of a gene aberrantly expressed in said disease or disorder.
9. The method of claim 8 , wherein said promoter is a globin promoter.
10. The method of claim 1 , wherein said looping factor consists of amino acids 1 to 200 of SEQ ID NO: 1.
11. The method of claim 1 , wherein said looping factor is an N-terminal fragment of Ldb1, wherein said N-terminal fragment of Ldb1 comprises amino acids 1 to 200 of SEQ ID NO: 1.
12. The method of claim 1 , wherein said method comprises administering a first and second nucleic acid molecule,
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 LIM domain binding 1 (Ldb1) or a fragment thereof, and wherein said first DNA binding domain specifically binds to a target sequence in the promoter of a 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 LIM domain binding 1 (Ldb1) or a fragment thereof, and wherein said second DNA binding domain specifically binds to a target sequence in the locus control region of said gene of interest.