NON-HUMAN ANIMALS WITH MODIFIED IMMUNOGLOBULIN HEAVY CHAIN SEQUENCES

摘要

Non-human animals, e.g., mammals, e.g., mice or rats, are provided comprising an immunoglobulin heavy chain locus that comprises a rearranged human immunoglobulin heavy chain variable region nucleotide sequence. The rearranged human immunoglobulin heavy chain variable region nucleotide sequence may be operably linked to a heavy or light chain constant region nucleic acid sequence. Also described are genetically modified non-human animals comprising an immunoglobulin light chain locus comprising one or more but less than the wild type number of human immunoglobulin light chain variable region gene segments, which may be operably linked to a light chain constant region nucleic acid sequence. Also provided are methods for obtaining nucleic acid sequences that encode immunoglobulin light chain variable domains capable of binding an antigen in the absence of a heavy chain.

主权项

1. A method of producing a nucleic acid that encodes a human immunoglobulin light chain variable domain comprising:
obtaining a nucleic acid comprising a sequence encoding the human immunoglobulin light chain variable domain from a lymphocyte of a rodent, or a hybridoma produced from the lymphocyte; wherein the rodent comprises in its germline genome
(i) at an endogenous immunoglobulin heavy chain locus a rearranged human immunoglobulin heavy chain variable region nucleotide sequence operably linked to an endogenous heavy chain constant region gene sequence, wherein the rearranged heavy chain variable region nucleotide sequence encodes the sequence of VH3-23/X1X2/4, wherein X1 is any amino acid, and X2 is any amino acid, and(ii) an immunoglobulin light chain locus comprising at least one unrearranged human immunoglobulin light chain variable gene segment (VL) and at least one unrearranged human immunoglobulin light chain J gene segment (JL) operably linked to an immunoglobulin light chain constant region gene sequence; wherein the lymphocyte expresses an immunoglobulin heavy chain variable domain that is (a) derived from the rearranged human immunoglobulin heavy chain variable region VH3-23/X1X2/JH gene linked to the endogenous heavy chain constant region gene sequence and (b) cognate with the human immunoglobulin light chain variable domain