Modular DNA-binding domains and methods of use
OwnerUlla Bonas, Jens Boch +2inventors
InventorsUlla Bonas, Jens Boch, Sebastian Schornack, Thomas Lahaye
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Abstract
The present invention refers to methods for selectively recognizing a base pair in a DNA sequence by a polypeptide, to modified polypeptides which specifically recognize one or more base pairs in a DNA sequence and, to DNA which is modified so that it can be specifically recognized by a polypeptide and to uses of the polypeptide and DNA in specific DNA targeting as well as to methods of modulating expression of target genes in a cell.
Claims17
A nucleic acid molecule encoding a protein comprising a DNA-binding domain and an additional domain,
- wherein the DNA-binding domain comprises a plurality of contiguous repeat units derived from a transcription activator-like (TAL) effector,
- wherein the additional domain is heterologous to a TAL effector and the additional domain has catalytic activity and is capable of modifying DNA, and
- wherein the protein is capable of recognizing a predetermined nucleotide sequence in a cell.
A nucleic acid molecule encoding a protein comprising a DNA-binding domain and an additional domain,
- wherein the DNA-binding domain comprises a plurality of contiguous repeat units derived from a transcription activator-like (TAL) effector,
- wherein the additional domain is heterologous to a TAL effector and the additional domain comprises a polypeptide or functional part or domain thereof selected from the group consisting of:
- a transcription repressor, a resistance-mediating protein, a nuclease, a topoisomerase, a ligase, an integrase, a recombinase, a resolvase, a methylase, an acetylase, a demethylase, and a deacetylase, and wherein the protein is capable of recognizing a predetermined nucleotide sequence in a cell.
A protein comprising a DNA-binding domain and an additional domain,
- wherein the DNA-binding domain comprises a plurality of contiguous repeat units derived from a transcription activator-like (TAL) effector,
- wherein the additional domain is heterologous to a TAL effector and the additional domain is a catalytic domain comprising an activity that is capable of modifying DNA, and
- wherein the protein is capable of recognizing a predetermined nucleotide sequence in a cell.
A protein comprising a DNA-binding domain and an additional domain,
- wherein the DNA-binding domain comprises a plurality of contiguous repeat units derived from a transcription activator-like (TAL) effector,
- wherein the additional domain is heterologous to a TAL effector and the additional domain comprises a polypeptide or functional part or domain thereof selected from the group consisting of:
- a transcription repressor, a resistance-mediating protein, a nuclease, a topoisomerase, a ligase, an integrase, a recombinase, a resolvase, a methylase, an acetylase, a demethylase, and a deacetylase, and
- wherein the protein is capable of recognizing a predetermined nucleotide sequence in a cell.
A method for producing a nucleic acid molecule encoding a protein comprising an engineered transcription activator-like (TAL) effector repeat domain, the method comprising:
- (a) combining nucleic acid segments encoding portions of a TAL effector repeat domain in an order based on the sequence of base pairs present in a predetermined target DNA sequence to produce a nucleotide sequence encoding an engineered TAL effector repeat domain capable of selectively binding the predetermined target DNA sequence, and
- (b) ligating the nucleic acid segments to produce a nucleic acid molecule encoding the protein comprising the engineered TAL effector repeat domain,
- wherein the protein is capable of binding selectively the predetermined target DNA sequence.
Dependencies are inferred from the claim text.