Regeneron Pharmaceuticals, Inc.

Corey Momont

Andrew J Murphy

Lynn Macdonald

John R McWhirter

David M Valenzuela

Ka-Man Venus Lai

Jose F Rojas

Gregg S Warshaw

Chris Schoenherr

Caitlin Montagna

Caitlin L Goshert

Naxin Tu

Vera Voronina

John Mcwhirter

Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends.

Nuclease-mediated DNA assembly

Provided herein are methods and compositions related to the in vivo testing of therapeutic agents comprising a human Fc in genetically modified rodents (e.g., the testing of the pharmacokinetic and/or pharmacodynamic properties of such a therapeutic agent in genetically modified rodents). In some embodiments the genetically modified rodents express antibodies comprising a human Fc (e.g., a human IgG1 Fc, a human IgG4 Fc). In some embodiments, the rodents express fully human antibodies (i.e., antibodies having human heavy chains and human light (&#x3b3; or &#x3ba;) chains). In certain embodiments the genetically modified rodents comprise one or more Fc receptors with a human extracellular domain (e.g., a Neonatal Fc Receptor (FcRn), a &#x3b2;-2-microglobulin polypeptide (&#x3b2;2M), a Fc &#x3b5; receptor 1&#x3b1; (Fc&#x3b5;R1&#x3b1;), a Fc &#x3b3; receptor 1 alpha (Fc&#x3b3;R1a), a Fc gamma receptor 2a (Fc&#x3b3;R2a), a Fc gamma receptor 2b (Fc&#x3b3;R2b), a Fc gamma receptor 3a (Fc&#x3b3;R3a), a Fc gamma receptor 3b (Fc&#x3b3;R3b), a Fc gamma receptor 2c (Fc&#x3b3;R2c)). The transmembrane and cytoplasmic domain of such receptors can be human or non-human (e.g., rodent).

Genetically modified mouse with humanized immunoglobulin heavy chain constant region genes and method of using

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New York, NY, United States of America

Corey Momont