US2011151539A1PendingUtilityA1

Use of meganucleases for inducing homologous recombination ex vivo and in toto in vertebrate somatic tissues and application thereof

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Assignee: ARNOULD SYLVAINPriority: Jan 28, 2003Filed: Feb 4, 2011Published: Jun 23, 2011
Est. expiryJan 28, 2023(expired)· nominal 20-yr term from priority
Inventors:Sylvain ArnouldSylvia BruneauJean-Pierre CabaniolsPatrick ChamesAndre ChoulikaPhilippe DuchateauJean-Charles EpinatAgnes GoubleEmmanuel LacroixFrederic PaquesChristophe Perez-MichautJulianne SmithDavid Sourdive
A61P 43/00A61P 31/18A61P 35/00A61P 35/02A61P 31/20A61P 31/12A61P 31/00C12N 2800/80C12N 15/8509C07K 2319/81C12Y 301/00C07K 14/435A61K 38/1709C12N 2840/44C12N 2730/10143C12N 2840/20C12N 15/86C12N 2830/002A61K 38/00A01K 2267/0337A01K 2267/03C12N 7/00A01K 67/0278C12Y 301/21004C12N 9/22A61K 48/0058C12N 2799/022A61K 48/00C12N 2830/55C12N 15/90C12N 15/1058A01K 2217/05A01K 2217/00A01K 2207/15C12N 15/907A01K 2227/105A61K 38/465A01K 67/0275
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Claims

Abstract

A single chain homing endonuclease, comprising a first variant of I-CreI having the amino acid sequence of accession number pdb 1g9y and a second variant of I-CreI variant having the amino acid sequence of accession number pdb 1g9y in a single polypeptide.

Claims

exact text as granted — not AI-modified
1 . A single chain homing endonuclease, comprising a first variant of I-CreI having the amino acid sequence of SEQ ID NO:23 and a second variant of I-CreI having the amino acid sequence of SEQ ID NO:23 in a single polypeptide, wherein
 said first variant of I-CreI contains the following substitutions: one to six substitutions in the sequence of amino acid residues 26 to 40 and one to three substitutions in the sequence of amino acid residues 44 to 77, wherein in said first variant of I-CreI there is a substitution at positions 33 and 68, and   independently, said second variant of I-CreI contains the following substitutions: four to six substitutions in the sequence of amino acid residues 26 to 40 and two to three substitutions in the sequence of amino acid residues 44 to 77, wherein in said second variant of I-CreI there is a substitution at positions 26, 30, 32, 38, 44, 68 and 77 wherein the endonuclease binds and cleaves DNA.   
     
     
         2 . The single chain homing endonuclease according to  claim 1 , wherein said second variant of I-CreI further contains a substitution at positions 40. 
     
     
         3 . The single chain homing endonuclease according to  claim 1 , wherein said first variant of I-CreI has a substitution at positions 24, 33 and 68. 
     
     
         4 . The single chain homing endonuclease according to  claim 1 , wherein said first variant of I-CreI has a substitution at positions 24, 33, 68 and 77. 
     
     
         5 . The single chain homing endonuclease according to  claim 1  wherein said second variant of I-CreI has a substitution at position 42.

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