Regeneron Pharmaceuticals, Inc.

Alexion Pharmaceuticals, Inc.

Mixing and Mass Transfer Technologies, LLC

Regeneran Pharmaceuticals, Inc.

The Penn State Research Foundation

General Signal Corporation

Other

Duke University

John R McWhirter

Andrew J Murphy

Lynn Macdonald

Sean Stevens

Cagan Gurer

Katherine S Bowdish

Vera Voronina

Faith Harris

Karolina Meagher

Prakash G Balan

Naxin Tu

Anke Kretz-Rommel

Toshiaki Maruyama

Corey Momont

David M Valenzuela

Ka-Man Venus Lai

Jose F Rojas

Gregg S Warshaw

Chunguang Guo

Chris Schoenherr

Margaret Karow

Joel H Martin

Yingzi Xue

Natasha Levenkova

Caitlin Montagna

Bradley A Swope

Paul M Kubera

George D Yancopoulos

David Frendewey

Aris I Baras

Israel Lowy

Brian Zambrowicz

Shana Frederickson

Matthew Koss

Marine Prissette

Garnett Kelsoe

David R Buckler

Bradley S Dominik

Barton F Haynes

Arijit Bhowmick

Mathieu Desclaux

Claudia Racioppi

Caitlin L Goshert

Ying Hu

Alexander O Mujica

Stanley J Wiegand

Jingtai Cao

Karolina A Hosiawa

Adrianna Latuszek

Ying-Chi Lin

Maruyama Toshiaki

James L Stanton

Non-human animals (and/or non-human cells) and methods of using the same are provided, which non-human animals (and/or non-human cells) have a genome comprising human antibody-encoding sequences (i.e., immunoglobulin genes). Non-human animals described herein express antibodies that contain immunoglobulin (Ig) light chains characterized by the presence of human Vλ domains. Non-human animals provided herein are, in some embodiments, characterized by expression of antibodies that contain human Vλ light chains that are encoded by human Igλ light chain-encoding sequences inserted into an endogenous Igκ light chain locus of said non-human animals. Methods for producing antibodies from non-human animals are also provided, which antibodies contain human variable regions and mouse constant regions.

Immunoglobulin lambda light chain and uses thereof

Provided herein are methods and compositions related to non-human animals that express exogenous Terminal Deoxynucleotidyltransferase (TdT).

Non-human animals expressing exogenous terminal deoxynucleotidyltransferase

BANF1, PPP2CA, and ANKLE2 were identified as genes that promote tau aggregation when disrupted. Improved tauopathy models such as cells, tissues, or animals having mutations in or inhibition of expression of BANF1 and/or PPP2CA and/or ANKLE2 are provided. Methods of using such improved tauopathy models for assessing therapeutic candidates for the treatment of a tauopathy, methods of making the improved tauopathy models, and methods of accelerating or exacerbating tau aggregation in a tauopathy model are also provided.

Models of tauopathy

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

Mice, embryos, cells, and tissues having a restricted immunoglobulin heavy chain locus and an ectopic sequence encoding one or more ADAM6 proteins are provided. In various embodiments, mice are described that have humanized endogenous immunoglobulin heavy chain loci and are capable of expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof that is functional in a male mouse. Mice, embryos, cells, and tissues having an immunoglobulin heavy chain locus characterized by a single human Vgene segment, a plurality of human Dgene segments and a plurality of human Jgene segments and capable expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof are also provided.

Humanized non-human animals with restricted immunoglobulin heavy chain loci

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

The invention provides a genetically modified non-human animal that comprises in its genome unrearranged T cell receptor variable gene loci, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified non-human animal and methods of making the same. Various methods of using the genetically modified non-human animal are also provided.

Genetically modified T cell receptor mice

Mice, tissues, cells, and genetic material are provided that comprise a humanized heavy chain immunoglobulin locus, a humanized light chain locus that expresses a universal light chain, and a gene encoding an ADAM6 or ortholog or homolog or functional fragment thereof. Mice are provided that express humanized heavy chains comprising human variable domains, and that express humanized light chains comprising human variable domains wherein the light chains are derived from no more than one, or no more than two, light chain V and J or rearranged V/J sequences. Fertile male mice that express antibodies with universal light chains and humanized heavy chains are provided. Methods and compositions for making bispecific binding proteins are provided.

Humanized universal light chain mice

Mice having a restricted immunoglobulin heavy chain locus are provided, wherein the locus is characterized by a single polymorphic human Vgene segment, a plurality of human Dgene segments and a plurality of Jgene segments. Methods for making antibody sequences that bind an antigen (e.g., a viral antigen) are provided, comprising immunizing a mouse with an antigen of interest, wherein the mouse comprises a single human Vgene segment, a plurality of human Dgene segments and a plurality of Jgene segments, at the endogenous immunoglobulin heavy chain locus.

Restricted immunoglobulin heavy chain mice

Genetically modified non-human animals are provided that express an immunoglobulin variable domain that comprises at least one histidine, wherein the at least one histidine is encoded by a substitution of a non-histidine codon in the germline of the animal with a histidine codon, or the insertion of a histidine codon in a germline immunoglobulin nucleic acid sequence. Immunoglobulin genes comprising histidines in one or more CDRs, in an N-terminal region, and/or in a loop 4 region are also provided. Immunoglobulin variable domains comprising one or more histidines (e.g., histidine clusters) substituted for non-antigen-binding non-histidine residues. Non-human animals that are progeny of animals comprising modified heavy chain variable loci (V, D, J segments), modified light chain variable loci (V, J segments), and rearranged germline light chain genes (VJ sequences) are also provided. Non-human animals that make immunoglobulin domains that bind antigens in a pH-sensitive manner are provided.

Non-human animals expressing pH-sensitive immunoglobulin sequences

A genetically modified non-human animal is provided, wherein the non-human animal expresses an antibody repertoire capable of pH dependent binding to antigens upon immunization. A genetically modified non-human animal is provided that expresses a single light chain variable domain derived from a single rearranged light chain variable region gene in the germline of the non-human animal, wherein the single rearranged light chain variable region gene comprises a substitution of at least one non-histidine encoding codon with a histidine encoding codon. Methods of making non-human animals that express antibodies comprising a histidine-containing universal light chain are provided.

Histidine engineered light chain antibodies and genetically modified non-human animals for generating same

Non-human animals (and/or non-human cells) and methods of using the same are provided, which non-human animals (and/or non-human cells) have a genome comprising human antibody-encoding sequences (i.e., immunoglobulin genes). Non-human animals described herein express antibodies that contain immunoglobulin (Ig) light chains characterized by the presence of human V&#x3bb; domains. Non-human animals provided herein are, in some embodiments, characterized by expression of antibodies that contain human V&#x3bb; light chains that are encoded by human Ig&#x3bb; light chain-encoding sequences inserted into an endogenous Ig&#x3ba; light chain locus of said non-human animals. Methods for producing antibodies from non-human animals are also provided, which antibodies contain human variable regions and mouse constant regions.

Mouse having an engineered immunoglobulin lambda light chain

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided.

Mice expressing a limited immunoglobulin light chain repertoire

Growing community of inventors

Boalsburg, PA, United States of America

John R McWhirter