Raif S Geha

US Patent:

6391548, May 21, 2002

Filed:

Mar 23, 1999

Appl. No.:

09/274383

Inventors:

John C. Bauer - San Diego CA
Dowain A. Wright - Cambridge MA
Jeffrey Carl Braman - Carlsbad CA
Raif S. Geha - Belmont MA

Assignee:

Stratagene - La Jolla CA
Childrens Medical Center Corp. - Longwood MA

International Classification:

C12Q 168

US Classification:

435 6, 435 912

Abstract:

The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells.

US Patent:

6635446, Oct 21, 2003

Filed:

Jun 22, 2000

Appl. No.:

09/599287

Inventors:

Narayanaswamy Ramesh - Wayland MA
Ines M. Anton - Brookline MA
John H. Hartwig - Jamaica Plain MA
Raif S. Geha - Belmont MA

Assignee:

The Childrens Medical Center Corporation - Boston MA

International Classification:

C12P 2106

US Classification:

435 691, 435325, 4352523, 4353201, 536 235, 530395

Abstract:

Described herein is a novel gene and its product, WIP, which associates with WASP. The subject invention relates to the isolated WIP gene or cDNA (see FIGS. A- B); nucleic acid probes, which can be fragments of the WIP gene or WIP cDNA or full-length; nucleic acid primers, which are fragments of WIP cDNA or the WIP gene; methods of assessing cells (e. g. , for diagnostic purposes) for the presence of WIP DNA, (e. g. , wildtype or mutated) or for the absence or occurrence of a reduced level of WIP DNA; WIP mRNA; WIP or WIP fragments, such as those which are useful to generate antibodies which bind WIP; and antibodies which bind WIP. Also the subject of this invention are methods of treating conditions in which WIP and/or WASP DNA or protein is deficient (in quantity) and/or defective (e. g. , mutated/altered) such that an individual is adversely affected (e. g. , has Wiskott-Aldrich Syndrome); methods of altering or regulating WASP and its functions; and methods of altering actin content, actin polymerization or both in cells, such as human lymphoid cells (e. g.

US Patent:

6713285, Mar 30, 2004

Filed:

May 10, 2002

Appl. No.:

10/144652

Inventors:

John C. Bauer - San Diego CA
Dowain A. Wright - Cambridge MA
Jeffrey Carl Braman - Carlsbad CA
Raif S. Geha - Belmont MA

Assignee:

Stratagene - La Jolla CA
Childrens Medical Center Corporation - Boston MA

International Classification:

C12P 1934

US Classification:

435 912

Abstract:

The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells.

US Patent:

6927318, Aug 9, 2005

Filed:

Feb 19, 2002

Appl. No.:

10/078547

Inventors:

Narayanaswamy Ramesh - Wayland MA, US
Miguel A. de la Fuente - Boston MA, US
Ines M. Anton - Turin, IT
Raif S. Geha - Belmont MA, US

Assignee:

The Children's Medical Center Corporation - Boston MA

International Classification:

C12N015/00
C12N005/00
A01K067/00
A01K067/027

US Classification:

800 21, 800 13, 800 18, 800 22, 800 25, 435325

Abstract:

Described herein is a novel gene and its product, WIP, which associates with WASP. The subject invention relates to the isolated WIP gene or cDNA and transgenic mammals that have the WIP gene disrupted in their genome. Also the subject of this invention are methods of treating conditions or diseases in which WIP and/or WASP DNA or protein is deficient and/or defective, for example, mutated or altered, such that an individual is adversely affected. Also described are methods of altering or regulating WIP and its functions in a mammal or in a cell of a mammal, for example in a lymphocyte. A further subject of this invention is an assay to identify drugs which alter the activity of WIP or expression of WIP DNA.

US Patent:

7132265, Nov 7, 2006

Filed:

Aug 26, 2002

Appl. No.:

10/229390

Inventors:

John C. Bauer - San Diego CA, US
Dowain A. Wright - Dunwoody GA, US
Jeffrey Carl Braman - Carlsbad CA, US
Raif S. Geha - Belmont MA, US

Assignee:

Stratagene California - La Jolla CA
Children's Medical Center Corporation - Boston MA

International Classification:

C12P 19/34

US Classification:

435 911, 435 912

Abstract:

The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells.

US Patent:

7176004, Feb 13, 2007

Filed:

Mar 25, 2004

Appl. No.:

10/811062

Inventors:

John C. Bauer - San Diego CA, US
Dowain A. Wright - Cambridge MA, US
Jeffrey Carl Braman - Carlsbad CA, US
Raif S. Geha - Belmont MA, US

Assignee:

Stratagene California - La Jolla CA
Children's Medical Center Corporation - Boston MA

International Classification:

C12P 19/34

US Classification:

435 912

Abstract:

The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells.

US Patent:

7402725, Jul 22, 2008

Filed:

May 20, 2005

Appl. No.:

11/134068

Inventors:

Narayanaswamy Ramesh - Wayland MA, US
Miguel A. de la Fuente - Boston MA, US
Ines M. Anton - Turin, IT
Raif S. Geha - Belmont MA, US

Assignee:

The Children's Medical Center Corporation - Boston MA

International Classification:

G01N 33/00
A01K 60/027
A01N 61/00
A61K 31/00

US Classification:

800 3, 800 14, 800 18, 514 1

Abstract:

Described herein is a novel gene and its product, WIP, which associates with WASP. The subject invention relates to the isolated WIP gene or cDNA and transgenic mammals that have the WIP gene disrupted in their genome. Also the subject of this invention are methods of treating conditions or diseases in which WIP and/or WASP DNA or protein is deficient and/or defective, for example, mutated or altered, such that an individual is adversely affected. Also described are methods of altering or regulating WIP and its functions in a mammal or in a cell of a mammal, for example in a lymphocyte. A further subject of this invention is an assay to identify drugs which alter the activity of WIP or expression of WIP DNA.

US Patent:

20070065926, Mar 22, 2007

Filed:

Sep 6, 2006

Appl. No.:

11/517188

Inventors:

John Bauer - San Diego CA, US
Dowain Wright - Cambridge MA, US
Jeffrey Braman - Carlsbad CA, US
Raif Geha - Belmont MA, US

International Classification:

C12P 19/34

US Classification:

435091200, 435455000

Abstract:

The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed. The double-stranded circular DNA intermediates is transformed in suitable competent host cells and closed circular double-stranded DNA corresponding to the parental template molecules, but containing the desired mutation or mutations of interest, may be conveniently recovered from the transformed cells. The invention also provide kits for site-directed mutagenesis in accordance with methods of the present invention.

Author:

Raif S. Geha

ISBN #:

0323018025

Author:

Raif S. Geha

ISBN #:

0815321740

Author:

Raif S. Geha

ISBN #:

0815333633

Author:

Raif S. Geha

ISBN #:

0815341024

Author:

Raif S. Geha

ISBN #:

0815340508