Nucleotide sequence encoding the enzyme I-SceI and the uses thereof
AssigneesInstitut Pasteur, Universite Pierre et Marie Curie
InventorsBernard Dujon, Andre Choulika, Arnaud Perrin, Jean-Francois Nicolas
Life of this patent
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Abstract
An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.
Claims34
1-22. (canceled)
A method for inducing at least one site directed double-stranded break in the DNA of an organism comprising:
- (a) providing an isolated cell of said organism comprising at least one endonuclease recognition site comprising 18 base pairs at a location in the DNA of the cell,
- (b) providing said endonuclease to said cell by genetically modifying the cell with a nucleic acid comprising said endonuclease or by introducing said endonuclease protein into the cell such that the endonuclease cleaves said endonuclease site at the location in the DNA of the cell.
A method for deleting a viral nucleic acid sequence from a target DNA in a cell comprising:
- providing a cell containing a viral nucleic acid sequence in said target DNA with at least one Group I intron encoded endonuclease sites within said viral nucleic acid sequence and
- expressing the Group I intron encoded endonuclease in the cell by genetically modifying the cell with a nucleic acid comprising said endonuclease or introducing said endonuclease protein into the cell such that the endonuclease cleaves the Group I intron encoded endonuclease site in the viral sequence,
- wherein a nucleic acid sequence is formed in the cell resulting a deletion of a viral nucleic acid sequence.
A method for deleting a viral nucleic acid sequence in a target nucleic acid sequence in an isolated cell, wherein said method comprises:
- providing an isolated cell containing a viral nucleic acid sequence in said target DNA with a Group I intron encoded endonuclease site within said viral nucleic acid sequence;
- expressing the Group I intron encoded endonuclease in the cell by genetically modifying the cell with a nucleic acid comprising said Group I intron encoded endonuclease or introducing said Group I intron encoded endonuclease protein into the cell;
- cleaving the Group I intron encoded endonuclease site with the endonuclease;
- and
- forming in the cell a nucleic acid sequence having a deletion of a viral nucleic acid sequence proximal to the Group I intron encoded endonuclease site.
A method for deleting a nucleic acid sequence from a target DNA in a cell comprising:
- providing an isolated cell containing a target DNA with two Group I intron encoded endonuclease sites within said target nucleic acid sequence;
- and
- expressing the Group I intron encoded endonuclease in the cell by genetically modifying the cell with a nucleic acid comprising said endonuclease or introducing said endonuclease protein into the cell such that the endonuclease cleaves the two Group I intron encoded endonuclease sites in the target nucleic acid sequence;
- wherein a nucleic acid sequence is formed in the cell having a deletion of a nucleic acid sequence between the two Group I intron encoded endonuclease sites.
A method for deleting a nucleic acid sequence in a target nucleic acid sequence in an isolated cell, wherein said method comprises:
- providing an isolated cell containing a Group I intron encoded endonuclease site within said target nucleic acid sequence;
- expressing the Group I intron encoded endonuclease in the cell by genetically modifying the cell with a nucleic acid comprising said Group I intron encoded endonuclease or introducing said Group I intron encoded endonuclease protein into the cell;
- cleaving the Group I intron encoded endonuclease site with the endonuclease;
- and
- forming in the cell a nucleic acid sequence having a deletion of a nucleic acid sequence proximal to the Group I intron encoded endonuclease site.
Dependencies are inferred from the claim text.