David J Heslin

US Patent:

20110041196, Feb 17, 2011

Filed:

Aug 13, 2010

Appl. No.:

12/856126

Inventors:

David FRENDEWEY - New York NY, US
David Jonathan HESLIN - Closter NJ, US
Ka-Man Venus LAI - Tarrytown NY, US
David M. VALENZUELA - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

C12N 15/87
C12N 15/85
C12N 5/10

US Classification:

800 21, 4353201, 435354

Abstract:

Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

US Patent:

20130095565, Apr 18, 2013

Filed:

Dec 3, 2012

Appl. No.:

13/692764

Inventors:

David Jonathan Heslin - Closter NJ, US
Ka-Man Venus Lai - Tarrytown NY, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

C12N 15/90

US Classification:

435354, 4353201

Abstract:

Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

US Patent:

20210130846, May 6, 2021

Filed:

Dec 10, 2020

Appl. No.:

17/117661

Inventors:

- Tarrytown NY, US
Wojtek Auerbach - Ridgewood NJ, US
David Heslin - Closter NJ, US
David Frendewey - New York NY, US
Ka-Man Venus Lai - Seattle WA, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

C12N 15/85
A01K 67/027
A61D 19/04
C12N 5/0735
A01K 67/02
C12N 15/877

Abstract:

Compositions and methods are provided for making rat pluripotent and totipotent cells, including rat embryonic stem (ES) cells. Compositions and methods for improving efficiency or frequency of germline transmission of genetic modifications in rats are provided. Such methods and compositions comprise an in vitro culture comprising a feeder cell layer and a population of rat ES cells or a rat ES cell line, wherein the in vitro culture conditions maintain pluripotency of the ES cell and comprises a media having mouse leukemia inhibitory factor (LIF) or an active variant or fragment thereof. Various methods of establishing such rat ES cell lines are further provided. Methods of selecting genetically modified rat ES cells are also provided, along with various methods to generate a transgenic rat from the genetically modified rat ES cells provided herein. Various kits and articles of manufacture are further provided.

US Patent:

20190316149, Oct 17, 2019

Filed:

May 2, 2019

Appl. No.:

16/401539

Inventors:

- Tarrytown NY, US
Wojtek Auerbach - Ridgewood NJ, US
David Heslin - Closter NJ, US
David Frendewey - New York NY, US
Ka-Man Venus Lai - San Francisco CA, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

C12N 15/85
A61D 19/04
A01K 67/027
C12N 5/0735
C12N 15/877
A01K 67/02

Abstract:

Compositions and methods are provided for making rat pluripotent and totipotent cells, including rat embryonic stem (ES) cells. Compositions and methods for improving efficiency or frequency of germline transmission of genetic modifications in rats are provided. Such methods and compositions comprise an in vitro culture comprising a feeder cell layer and a population of rat ES cells or a rat ES cell line, wherein the in vitro culture conditions maintain pluripotency of the ES cell and comprises a media having mouse leukemia inhibitory factor (LIF) or an active variant or fragment thereof. Various methods of establishing such rat ES cell lines are further provided. Methods of selecting genetically modified rat ES cells are also provided, along with various methods to generate a transgenic rat from the genetically modified rat ES cells provided herein. Various kits and articles of manufacture are further provided.

US Patent:

20170037429, Feb 9, 2017

Filed:

Aug 19, 2016

Appl. No.:

15/242025

Inventors:

- Tarrytown NY, US
Wojtek Auerbach - Ridgewood NJ, US
David Heslin - Closter NJ, US
David Frendewey - New York NY, US
Ka-Man Venus Lai - Tarrytown NY, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

C12N 15/85
A01K 67/02
C12N 15/877
A01K 67/027

Abstract:

Compositions and methods are provided for making rat pluripotent and totipotent cells, including rat embryonic stem (ES) cells. Compositions and methods for improving efficiency or frequency of germline transmission of genetic modifications in rats are provided. Such methods and compositions comprise an in vitro culture comprising a feeder cell layer and a population of rat ES cells or a rat ES cell line, wherein the in vitro culture conditions maintain pluripotency of the ES cell and comprises a media having mouse leukemia inhibitory factor (LIF) or an active variant or fragment thereof. Various methods of establishing such rat ES cell lines are further provided. Methods of selecting genetically modified rat ES cells are also provided, along with various methods to generate a transgenic rat from the genetically modified rat ES cells provided herein. Various kits and articles of manufacture are further provided.

US Patent:

20160150767, Jun 2, 2016

Filed:

Jan 7, 2016

Appl. No.:

14/990313

Inventors:

- Tarrytown NY, US
David Jonathan Heslin - Closter NJ, US
Ka-Man Venus Lai - Tarrytown NY, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

Regeneron Pharmaceuticals, Inc. - Tarrytown NY

International Classification:

A01K 67/027

Abstract:

Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

US Patent:

20160108360, Apr 21, 2016

Filed:

Oct 30, 2015

Appl. No.:

14/928134

Inventors:

- Tarrytown NY, US
Wojtek Auerbach - Ridgewood NJ, US
David Heslin - Closter NJ, US
David Frendewey - New York NY, US
Ka-Man Venus Lai - Tarrytown NY, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

REGENERON PHARMACEUTICALS, INC. - Tarrytown NY

International Classification:

C12N 5/0735

Abstract:

Compositions and methods are provided for making rat pluripotent and totipotent cells, including rat embryonic stem (ES) cells. Compositions and methods for improving efficiency or frequency of germline transmission of genetic modifications in rats are provided. Such methods and compositions comprise an in vitro culture comprising a feeder cell layer and a population of rat ES cells or a rat ES cell line, wherein the in vitro culture conditions maintain pluripotency of the ES cell and comprises a media having mouse leukemia inhibitory factor (LIF) or an active variant or fragment thereof. Various methods of establishing such rat ES cell lines are further provided. Methods of selecting genetically modified rat ES cells are also provided, along with various methods to generate a transgenic rat from the genetically modified rat ES cells provided herein. Various kits and articles of manufacture are further provided.

US Patent:

20150291978, Oct 15, 2015

Filed:

Jun 25, 2015

Appl. No.:

14/750318

Inventors:

- Tarrytown NY, US
David Jonathan Heslin - Closter NJ, US
Ka-Man Venus Lai - Tarrytown NY, US
David M. Valenzuela - Yorktown Heights NY, US

Assignee:

REGENERON PHARMACEUTICALS, INC. - Tarrytown NY

International Classification:

C12N 15/85
C12N 9/12

Abstract:

Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.