Tevi chimeric endonuclease and their preferential cleavage sites

1. A method to selectively cleave a target nucleic acid in a mammalian cell using a catalytic domain of I-TevI, said method comprising:

a) selecting a target nucleic acid sequence in a mammalian cell comprising a cleavage site having the motif CNNNGN selected from the group consisting of: SEQ ID NO: 2 to 55;

b) selecting or engineering a nucleic acid binding domain to specifically bind a recognition site adjacent to said cleavage site; wherein the engineered nucleic acid binding domain is an engineered Transcription Activator Like Effectors (TALE) binding domain comprising a plurality of TALE like repeat sequences, each repeat comprising a repeat variable dipeptide (RVD) specific to each nucleotide base of a TALE DNA binding site;

c) fusing said nucleic acid binding domain with said catalytic domain of I-TevI to obtain a TevI chimeric endonuclease,

wherein said TevI chimeric endonuclease is selected from the group consisting of: SEQ ID NO: 452 to 458; and

d) contacting said target nucleic acid with said chimeric endonuclease in said mammalian cell,

wherein said catalytic domain of I-TevI cleaves the target nucleic acid in the mammalian cell at said cleavage site.

2. The method of claim 1 , further comprising a step of introducing said cleavage site selected in step a) into said target nucleic acid in order to be cleaved by said TevI chimeric endonuclease.

3. The method of claim 1 , wherein said catalytic domain of I-TevI has the protein sequence of SEQ ID NO: 257.

4. The method of claim 1 , wherein said catalytic domain of I-TevI is selected from the group consisting of: SEQ ID NO: 268 and SEQ ID NO: 284.

5. The method of claim 1 , wherein said catalytic domain of I-TevI shares at least 80% protein sequence identity with a protein sequence selected from the group consisting of: SEQ ID NO: 268 and SEQ ID NO: 284.

6. The method of claim 1 , wherein said catalytic domain of I-TevI is fused to the N-terminal part of said nucleic acid binding domain.

7. The method of claim 1 , wherein said catalytic domain of I-TevI is fused to the C-terminal part of said nucleic acid binding domain.

8. The method of claim 1 , wherein said catalytic domain of I-TevI is fused to said nucleic acid binding domain by a peptide linker.

9. The method of claim 1 , wherein the TALE nucleic acid recognition site comprises a T nucleotide (T 0 ) located between 5 to 20 bp away from the G base in the motif CNNNGN of said I-TevI cleavage site.

10. The method of claim 1 , wherein the TALE nucleic acid recognition site comprises a T nucleotide (T 0 ) located between 6 to 12 bp away from the G base in the motif CNNNGN of said I-TevI cleavage site.

11. The method of claim 1 , wherein the TALE nucleic acid recognition site comprises a T nucleotide (T 0 ) located 10 bp away from the G base in the motif CNNNGN of said I-TevI cleavage site.

12. The method according to claim 1 , wherein said target nucleic acid sequence comprises a nucleotide A or T after the G base in the motif CNNNGN of said I-TevI cleavage site.

13. The method according to claim 1 , further comprising providing to the mammalian cell an exogenous nucleic acid comprising a sequence homologous to at least a portion of the target nucleic acid sequence, such that homologous recombination occurs between the target DNA sequence and the exogenous nucleic acid.

14. The method according to claim 1 , further comprising a step of providing and contacting an additional catalytic domain with the target nucleic acid sequence.

15. The method of claim 14 , wherein said additional catalytic domain is a DNA end-processing enzyme.

16. The method of claim 15 , wherein said DNA end-processing enzyme is Trex2.

17. The method of claim 15 , wherein said DNA processing enzyme is a single chain Trex2.