Lambda integrase mediated recombination in plants
AssigneeSyngenta Participations AG
InventorsJanet Louise Suttie, Mary-Dell Chilton, Qiudeng Que
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Abstract
The present disclosure provides methods for obtaining the targeted integration of a DNA molecule into the genome of a host cell using a recombinase. The methods disclosed herein can be used with a variety of host cells, including, for example, dicotyledonous and monocotyledonous plant cells. The present disclosure provides a method for effecting site-specific recombination of DNA within a plant cell, comprising: introducing into the plant cell a target nucleotide sequence comprising a first Int recognition site; introducing into the plant cell a donor nucleotide sequence comprising a second Int recognition site; and introducing into the plant cell an integrase or integrase complex.
Claims119
A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
- introducing into a plant cell a target sequence comprising a first Int recognition site capable of being recognized by a Lambda bacteriophage integrase or integrase complex;
- introducing into said plant cell a donor sequence comprising a second Int recognition site capable of being recognized by a Lambda bacteriophage integrase or integrase complex;
- and
- introducing into said plant cell a Lambda bacteriophage integrase or integrase complex, such that site-specific recombination of said donor sequence is obtained.
A method for manipulating a target sequence within a plant cell, comprising:
- introducing into a plant cell a target sequence comprising (a) a first Int recognition site capable of being recognized by a Lambda bacteriophage integrase or integrase complex and a second Int recognition site capable of being recognized by a Lambda bacteriophage integrase or integrase complex that are capable of recombining with each other and (b) a first nucleotide sequence that is situated between said first Int recognition site and said second Int recognition site;
- introducing into said plant cell a Lambda bacteriophage integrase or integrase complex;
- and
- identifying a recombination product comprising an altered target sequence.
A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
- introducing into a genome of a plant cell a target sequence comprising a first Int recognition site that comprises attL (SEQ ID NO:174);
- introducing into said plant cell a donor sequence comprising a second Int recognition site that comprises attR (SEQ ID NO:175);
- introducing into said plant cell a Lambda bacteriophage integrase or integrase complex;
- and
- identifying a recombination product within said genome of said plant cell obtained through sequence exchange between said target sequence and said donor sequence.
A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
- introducing into a genome of a plant cell a target sequence comprising attL (SEQ ID NO:174) as a first Int recognition site and comprising attL1 (SEQ ID NO:180) as a third Int recognition site;
- introducing into said plant cell a donor sequence comprising attR (SEQ ID NO:175) as a second Int recognition site and comprising attR1 (SEQ ID NO:181) as a fourth recognition site;
- introducing into said plant cell a Lambda bacteriophage integrase or integrase complex;
- and
- identifying a recombination product within said genome of said plant cell obtained through sequence exchange between said target sequence and said donor sequence.
Dependencies are inferred from the claim text.