David C Segal

US Patent:

20030232410, Dec 18, 2003

Filed:

Mar 20, 2003

Appl. No.:

10/395816

Inventors:

Monika Liljedahl - La Jolla CA, US
Simon Aspland - San Diego CA, US
David Segal - Tucson AZ, US

International Classification:

C12P021/02
C12N015/85
C12N005/06

US Classification:

435/069100, 435/455000, 435/325000

Abstract:

Embodiments relate to methods of generating a genetically modified cell. The methods can include providing a primary cell containing an endogenous chromosomal target DNA sequence in which it is desired to have homologous recombination occur. The methods also can include providing a zinc finger endonuclease (ZFE) that includes an endonuclease domain that cuts DNA, and a zinc finger domain that includes a plurality of zinc fingers that bind to a specific nucleotide sequence within the endogenous chromosomal target DNA in the primary cell. Further, the methods can include contacting the endogenous chromosomal target DNA sequence with the zinc finger endonuclease in the primary cell such that the zinc finger endonuclease cuts both strands of a nucleotide sequence within the endogenous chromosomal target DNA sequence in the primary cell, thereby enhancing the frequency of homologous recombination in the endogenous chromosomal target DNA sequence. The methods also include providing a nucleic acid comprising a sequence homologous to at least a portion of said endogenous chromosomal target DNA such that homologous recombination occurs between the endogenous chromosomal target DNA sequence and the nucleic acid.

US Patent:

20080209587, Aug 28, 2008

Filed:

Oct 17, 2007

Appl. No.:

11/975017

Inventors:

Monika Liljedahl - La Jolla CA, US
Simon Eric Aspland - San Diego CA, US
David J. Segal - Tucson AZ, US

International Classification:

C12N 15/82
A01H 5/00

US Classification:

800278, 800298

Abstract:

Embodiments relate to methods of generating a genetically modified cell. The methods can include providing a primary cell containing an endogenous chromosomal target DNA sequence in which it is desired to have homologous recombination occur. The methods also can include providing a zinc finger endonuclease (ZFE) that includes an endonuclease domain that cuts DNA, and a zinc finger domain that includes a plurality of zinc fingers that bind to a specific nucleotide sequence within the endogenous chromosomal target DNA in the primary cell. Further, the methods can include contacting the endogenous chromosomal target DNA sequence with the zinc finger endonuclease in the primary cell such that the zinc finger endonuclease cuts both strands of a nucleotide sequence within the endogenous chromosomal target DNA sequence in the primary cell, thereby enhancing the frequency of homologous recombination in the endogenous chromosomal target DNA sequence. The methods also include providing a nucleic acid comprising a sequence homologous to at least a portion of said endogenous chromosomal target DNA such that homologous recombination occurs between the endogenous chromosomal target DNA sequence and the nucleic acid.

US Patent:

20090305402, Dec 10, 2009

Filed:

Jun 11, 2009

Appl. No.:

12/456111

Inventors:

Monika Liljedahl - La Jolla CA, US
Simon Eric Aspland - San Diego CA, US
David J. Segal - Tucson AZ, US

International Classification:

C12N 5/10
C12N 5/04
C12N 5/06
C12N 5/08

US Classification:

435349, 435419, 435325, 435366, 435363, 435353, 435354

Abstract:

Described herein are methods of generating a genetically modified cell by providing a zinc finger endonuclease (ZFE) that includes an endonuclease domain that cuts DNA, and a zinc finger domain that includes a plurality of zinc fingers that bind to a specific nucleotide sequence within the endogenous chromosomal target DNA in the primary cell. Further, the methods can include contacting the endogenous chromosomal target DNA sequence with the zinc finger endonuclease in the primary cell such that the zinc finger endonuclease cuts both strands of a nucleotide sequence within the endogenous chromosomal target DNA sequence in the primary cell, thereby enhancing the frequency of homologous recombination in the endogenous chromosomal target DNA sequence.