TAL effector-mediated DNA modification
Materials and methods related to gene targeting (e.g., gene targeting with transcription activator-like effector nucleases; “TALENS”) are provided.
1. An isolated human cell comprising a nucleic acid encoding a transcription activator-like (TAL) effector endonuclease, the TAL effector endonuclease comprising:
(a) a FokI endonuclease domain that can cleave double stranded DNA, and
(b) a TAL effector domain comprising a plurality of TAL effector repeat sequences that, in combination, bind to a target DNA sequence in said human cell.
2. The isolated human cell of claim 1 , wherein the TAL effector domain comprises 15 or more DNA binding repeats.
3. The isolated human cell of claim 1 , wherein each TAL effector repeat sequence comprises a repeat variable-diresidue (RVD) that determines recognition of a base pair in the target DNA sequence, wherein each DNA binding repeat is responsible for recognizing one base pair in the target DNA sequence, and wherein the RVD comprises one or more of:
HD for recognizing C;
NG for recognizing T;
NI for recognizing A;
NN for recognizing G;
NS for recognizing A;
HG for recognizing T;
IG for recognizing T;
NK for recognizing G;
HA for recognizing C;
HI for recognizing C;
HN for recognizing G; and
NA for recognizing G.