Methods of transcription activator like effector assembly
US-10273271-B2 · Apr 30, 2019 · US
TAL-Tet1 fusion proteins and methods of use thereof
US10894950B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10894950-B2 |
| Application number | US-201815894004-A |
| Country | US |
| Kind code | B2 |
| Filing date | Feb 12, 2018 |
| Priority date | May 29, 2012 |
| Publication date | Jan 19, 2021 |
| Grant date | Jan 19, 2021 |
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Abstract
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Fusion proteins comprising a DNA binding domain, e.g., a TAL effector repeat array or zinc finger, and a catalytic domain comprising a sequence that catalyzes hydroxylation of methylated cytosines in DNA, and methods of use thereof.
First claim
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What is claimed is: 1. A nucleic acid encoding a fusion protein comprising an engineered transcription activator-like (TAL) effector repeat array or a zinc finger protein DNA-binding domain that is capable of specifically binding to a DNA sequence that is within 250 bp of a CpG diresidue in a mammalian cell, fused to a catalytic domain comprising a catalytic domain of Tet1 comprising amino acids 1580-2052 of SEQ ID NO: 2, and excluding amino acids 584-624 of SEQ ID NO:2, that is capable of catalyzing hydroxylation of methylated cytosines in DNA. 2. The nucleic acid of claim 1 , wherein the fusion protein further comprises a linker between the DNA binding domain and the catalytic domain. 3. The nucleic acid of claim 1 , wherein the fusion protein comprises a plurality of catalytic domains. 4. The nucleic acid of claim 1 , wherein the DNA-binding domain comprises a six-finger zinc finger protein. 5. The nucleic acid of claim 1 , wherein the catalytic domain consists of the amino acid sequence of SEQ ID NO:4. 6. A vector comprising the nucleic acid of claim 1 . 7. The vector of claim 6 , wherein the vector is a DNA plasmid, an adeno-associated virus, or a retrovirus. 8. A composition comprising the nucleic acid of claim 1 and a delivery vehicle. 9. The composition of claim 8 , wherein the delivery vehicle is a liposome. 10. A method of reducing methylation of a CpG diresidue in a mammalian cell, the method comprising contacting the cell with a nucleic acid encoding a fusion protein comprising an engineered transcription activator-like (TAL) effector repeat array or a zinc finger protein DNA-binding domain that is capable of specifically binding to a DNA sequence that is within 250 bp of a CpG diresidue in a mammalian cell, fused to a catalytic domain comprising a catalytic domain of Tet1 comprising amino acids 1580-2052 of SEQ ID NO: 2, and excluding amino acids 584-624 of SEQ ID NO:2, that is capable of catalyzing hydroxylation of methylated cytosines in DNA, and wherein the fusion protein reduces methylation of the CpG diresidue. 11. The method of claim 10 , wherein the cell is a human cell. 12. The method of claim 10 , wherein the DNA sequence is a gene selected from the group consisting of RASSF1A, MGMT, BRCA1, p16, CDH1, TIMP-3, p15, p73, DAPK, APC, and RELN. 13. The method of claim 10 , wherein the DNA sequence is within 100 bp of the CpG diresidue. 14. The method of claim 10 , wherein the DNA sequence is within 30 bp of the CpG diresidue.
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Classifications
containing a Zn-finger domain for DNA binding · CPC title
containing a DNA binding domain, e.g. Lacl or Tet-repressor · CPC title
- C12N9/0071Primary
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Drugs for disorders of the cardiovascular system · CPC title
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- What does patent US10894950B2 cover?
- Fusion proteins comprising a DNA binding domain, e.g., a TAL effector repeat array or zinc finger, and a catalytic domain comprising a sequence that catalyzes hydroxylation of methylated cytosines in DNA, and methods of use thereof.
- Who is the assignee on this patent?
- Massachusetts Gen Hospital
- What technology area does this patent fall under?
- Primary CPC classification C12N9/0071. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jan 19 2021 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 6 related publications on this page (citations in our corpus or others sharing the same primary CPC).