NON-HUMAN ANIMALS THAT MAKE SINGLE DOMAIN BINDING PROTEINS

摘要

Genetically modified non-human animals and methods and compositions for making and using them are provided, wherein the genetic modification comprises (a) a deletion in an immunoglobulin constant region CH1 gene (optionally a deletion in a hinge region) of a heavy chain constant region gene sequence, and (b) replacement of one or all endogenous VH, DH and JH gene segments with at least one unrearranged light chain variable (VL) gene segment and at least one unrearranged light chain joining (JL) gene segment capable of recombining to form a rearranged light chain variable region (VL/JL) nucleotide sequence operably linked to the heavy chain constant region gene sequence comprising a deletion in the CH1 gene and/or insertion of a genetically engineered single rearranged light chain, wherein the mouse is capable of expressing a functional IgM, single domain antigen binding proteins, e.g., VL-single domain binding proteins, and a genetically engineered rearranged light chain.

主权项

1. A genetically modified non-human animal comprising in its germline
(a) a deletion or inactivating mutation in a nucleotide sequence encoding a CH1 domain of at least one endogenous immunoglobulin heavy chain constant region gene at an endogenous immunoglobulin heavy chain locus, wherein the at least one endogenous immunoglobulin heavy chain constant region gene is IgG, IgA, IgE, IgD, or a combination thereof, and (b) either or both
(i) a nucleic acid sequence comprising at least one unrearranged immunoglobulin light chain variable region (VL) gene segment and at least one unrearranged immunoglobulin light chain joining (JL) gene segment, wherein the unrearranged VL and JL gene segments are capable of recombining to form a rearranged immunoglobulin light chain variable region (VL/JL) nucleotide sequence operably linked to the immunoglobulin heavy chain constant region gene comprising the deletion or inactivating mutation in the nucleotide sequence encoding the CH1 domain, and/or(ii) an immunoglobulin light chain locus that comprises a single rearranged immunoglobulin light chain variable region VL/JL gene sequence comprising VL and JL gene segment sequences, wherein the single rearranged immunoglobulin light chain variable region gene sequence is operably linked to an immunoglobulin light chain constant region gene sequence.