Lambda integrase mediated recombination in plants
First Claim
1. 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;
introducing into said plant cell a donor sequence comprising a second Int recognition site; and
introducing into said plant cell an integrase or integrase complex.
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Accused Products
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.
2 Citations
133 Claims
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1. A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
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introducing into a plant cell a target sequence comprising a first Int recognition site;
introducing into said plant cell a donor sequence comprising a second Int recognition site; and
introducing into said plant cell an integrase or integrase complex. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119)
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88. A method for manipulating a target sequence within a plant cell, comprising:
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introducing into a plant cell a target sequence comprising (a) a first Int recognition site and a second Int recognition site 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 an integrase or integrase complex; and
identifying a recombination product comprising an altered target sequence. - View Dependent Claims (89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99)
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100. A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
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introducing into a genome of a plant cell a target sequence comprising a first Int recognition site that comprises one of attL (SEQ ID NO;
174), attL1 (SEQ ID NO;
180), attL2 (SEQ ID NO;
182), attR (SEQ ID NO;
175), attR1 (SEQ ID NO;
181), attR2 (SEQ ID NO;
183), attL3, and attR3;
introducing into said plant cell a donor sequence comprising a second Int recognition site that comprises one of attL (SEQ ID NO;
174), attL1 (SEQ ID NO;
180), attL2 (SEQ ID NO;
182), attR (SEQ ID NO;
175), attR1 (SEQ ID NO;
181), attR2 (SEQ ID NO;
183), attL3, and attR3;
introducing into said plant cell an 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. - View Dependent Claims (101, 102, 103, 104)
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105. A method for obtaining site-specific recombination of DNA within a plant cell, comprising:
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introducing into a genome of a plant cell a target sequence comprising a first Int recognition site and a third Int recognition site, wherein each of said first Int recognition site and said third Int recognition site is independently selected and comprises one of attL (SEQ ID NO;
174), attL1 (SEQ ID NO;
180), attL2 (SEQ ID NO;
182), attR (SEQ ID NO;
175), attB1 (SEQ ID NO;
181), attR2 (SEQ ID NO;
183), attL3, and attR3;
introducing into said plant cell a donor sequence comprising a second Int recognition site and a fourth recognition site, wherein each of said second Int recognition site and said fourth Int recognition site is independently selected and comprises one of attL (SEQ ID NO;
174), attL1 (SEQ IDNO;
180), attL2 (SEQ ID NO;
.182). attR (SEQ ID NO;
175), attR1 (SEQ ID NO;
181), attR2 (SEQ ID NO;
183), attL3, and attR3;
introducing into said plant cell an 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. - View Dependent Claims (106, 107, 108, 109)
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- 120. An isolated nucleotide sequence comprising SEQ ID NO:
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132. An isolated nucleotide sequence comprising SEQ ID NO:
- 29 and SEQ ID NO;
37.
- 29 and SEQ ID NO;
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133. An isolated nucleotide sequence comprising SEQ ID NO:
- 43.
Specification