ENGINEERED TAL EFFECTOR PROTEINS WITH ENHANCED DNA TARGETING CAPACITY

摘要

The present invention provides compositions and methods for DNA targeting using TAL effectors and TAL effector based proteins, including but not limited to targeted gene regulation and targeted cleavage of cellular chromatin in a region of interest and/or homologous recombination at a predetermined site in cells. Compositions include fusion polypeptides comprising a TAL effector or a TAL effector binding domain in combination with other domains, including but not limited to a cleavage domain. The TAL effector binding domain includes modifications that increase activity of the same and also remove the constraints that the DNA target sequence be preceded by a thymine.

主权项

1. A method for modifying the genetic material of a cell, comprising:
(a) providing a cell containing a target DNA sequence; and (b) introducing a transcription activator-like (TAL) effector-DNA modifying enzyme (TALEN) into the cell, the TALEN comprising:
(i) a DNA modifying enzyme domain that can modify double stranded DNA, and(ii) a TAL effector domain comprising a plurality of TAL effector repeat sequences that, in combination, bind to a specific nucleotide sequence in the target DNA sequence, and further wherein said effector repeat sequences are such that the target DNA sequence does not require a 5′ thymine residue, and/or wherein said activity of TAL effector is increased.