Transcription activator-like effector nucleases (TALENs)

1. A polynucleotide sequence encoding a transcription activator like effector nuclease (TALEN), the TALEN comprising from an N-terminus to a C-terminus:

(i) a first segment of about 50 to about 200 amino acids in length, wherein the first segment comprises the amino acid sequence of SEQ ID NO: 1;

(ii) a TAL effector DNA-binding domain providing sequence-specific binding to a target nucleotide sequence, wherein the TAL effector DNA-binding domain comprises 12-31 TAL repeats and a C-terminal truncated TAL repeat;

(iii) a second segment of about 20 to 100 amino acids in length, wherein the first segment comprises the amino acid sequence of SEQ ID NO: 2; and

(iv) a modified FokI nuclease catalytic domain.

2. The polynucleotide sequence of claim 1 , wherein the polynucleotide is mRNA.

3. An expression cassette comprising a promoter and further comprising the polynucleotide of claim 1 .

4. The expression cassette of claim 3 , further comprising a coding sequence for a nuclear localization signal (NLS) and a polyadenylation signal sequence.

5. The expression cassette of claim 4 , wherein the NLS comprises an SV40 NLS.

6. The expression cassette of claim 5 , wherein the SV40 NLS comprises PKKKRKV (SEQ ID NO:649).

7. The expression cassette of claim 4 , wherein the expression cassette is the plasmid pCS2-TALENs-ELD or pCS2-TALENs-KKR.

8. A host cell comprising the expression cassette of claim 3 .

9. A method of producing the mRNA of claim 2 , comprising providing a polynucleotide sequence of claim 1 , wherein the polynucleotide sequence is DNA, under conditions permissible for mRNA transcription.

10. The polynucleotide of claim 1 , wherein the modified FokI nuclease catalytic domain is an obligate heterodimer.

11. The polynucleotide of claim 1 , wherein the modified FokI nuclease catalytic domain has the amino acid sequence of SEQ ID NOs:3 or 4.