Methods of transcription activator like effector assembly

Patent metadata

Field	Value
Publication number	US-11472849-B2
Application number	US-201916283380-A
Country	US
Kind code	B2
Filing date	Feb 22, 2019
Priority date	Jul 15, 2011
Publication date	Oct 18, 2022
Grant date	Oct 18, 2022

Abstract

The disclosure describes methods that include providing a first nucleic acid having a sequence encoding a first set comprising one or more transcription activator-like effector (TALE) repeat domains and/or one or more portions of one or more TALE repeat domains; contacting the first nucleic acid with a first enzyme, wherein the first enzyme creates a first ligatable end; providing a second nucleic acid having a sequence encoding a second set comprising one or more TALE repeat domains and/or one or more portions of one or more TALE repeat domains; contacting the second nucleic acid with a second enzyme, wherein the second enzyme creates a second ligatable end, and wherein the first and second ligatable ends are compatible; and ligating the first and second nucleic acids through the first and second ligatable ends to produce a first ligated nucleic acid, wherein the first ligated nucleic acid is linked to a solid support, and wherein the first ligated nucleic acid encodes a polypeptide comprising said first and second sets.

First claim

What is claimed is:

1. A composition comprising a plurality of nucleic acids encoding one or more transcription activator-like effector (TALE) repeat domains, wherein at least one nucleic acid is selected from the group consisting of SEQ ID NOs: 58-62.

2. The composition of claim 1 , further comprising one or more of:
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 63-67
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 68-72;
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 73-77;
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 78-82;
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 83-87; or
at least one nucleic acid selected from the group consisting of SEQ ID NOs: 88-92.

3. The composition of claim 2 , wherein the plurality of nucleic acids comprises each of SEQ ID NOs: 58-92.

4. The composition of claim 1 , wherein the nucleic acid encodes a functional TALE protein that is 33-35 amino acids.

5. The composition of claim 1 , wherein each nucleic acid of the composition is in a plasmid.

6. The composition of claim 5 , wherein each plasmid of the composition is in a cell.

7. The composition of claim 1 , wherein the at least one nucleic acid is linked to a solid support.

8. A plasmid comprising at least one nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs: 58-62.

9. The plasmid of claim 8 , wherein the plasmid is in a cell.

Assignees

• Massachusetts Gen Hospital

Inventors

• Joung J Keith
• Sander Jeffry D

Classifications

• C12P19/34

  Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title

• C12N15/66

  General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease · CPC title

• C07K2319/43

  containing a FLAG-tag · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C07K14/195 Primary

  from bacteria · CPC title