Non-human animals with modified immunoglobulin heavy chain sequences

Abstract

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.

1 .- 106 . (canceled) 
     
     
         107 . A nucleotide molecule comprising a rearranged human immunoglobulin heavy chain variable region nucleotide sequence that comprises a human germline heavy chain V H  gene segment and/or a human germline heavy chain J H  gene segment. 
     
     
         108 . The nucleotide molecule of  claim 107 , wherein
 (i) the human germline heavy chain V H  gene segment is selected from the group consisting of V H 1-2, V H 1-3, V H 1-8, V H 1-18, V H 1-24, V H 1-45, V H 1-46, V H 1-58, V H 1-69, V H 2-5, V H 2-26, V H 2-70, V H 3-7, V H 3-9, V H 3-11, V H 3-13, V H 3-15, V H 3-16, V H 3-20, V H 3- 21, V H 3-23, V H 3-30, V H 3-30-3, V H  3-30-5, V H 3-33, V H 3-35, V H 3-38, V H 3-43, V H 3-48, V H 3-49, V H 3-53, V H 3-64, V H 3-66, V H 3-72, V H 3-73, V H 3-74, V H 4-4, V H 4-28, V H 4-30-1, V H 4-30-2, V H 4-30-4, V H 4-31, V H 4-34, V H 4-39, V H 4-59, V H 4-61, V H 5-51, V H 6-1, V H 7-4-1, V H 7-81, and any polymorphic variant thereof.   and/or   (ii) the human germline heavy chain J H  gene segment is selected from the group consisting of J H 1, J H 2, J H 3, J H 4, J H 5, J H 6, and any polymorphic variant thereof   
     
     
         109 . The nucleotide molecule of  claim 107 , wherein the rearranged human immunoglobulin heavy chain variable region sequence comprises a human germline heavy chain V H  gene segment and a human germline heavy chain J H  gene segment. 
     
     
         110 . The nucleotide molecule of 107, wherein the rearranged human immunoglobulin heavy chain variable region sequence encodes the sequence of V H 3-23/X 1 X2/J H , wherein X 1  is any amino acid, and X2 is any amino acid. 
     
     
         111 . The nucleotide molecule of  claim 110 , wherein X 1  is Gly and X 2  is Tyr. 
     
     
         112 . The nucleotide molecule of  claim 111 , wherein J H  is selected from the group consisting of a human germline J H 1 gene segment, a human germline J H 2 gene segment, a human germline J H 3 gene segment, a human germline J H 4 gene segment, a human germline J H 5 gene segment, a human germline J H 6 gene segment, and a polymorphic variant thereof. 
     
     
         113 . The nucleotide molecule of  claim 107 , wherein the rearranged human immunoglobulin heavy chain variable region sequence encodes the sequence of human V H 3-23/GY/J H 4-4 (SEQ ID NO:137). 
     
     
         114 . The nucleotide molecule of  claim 107 , comprising the rearranged human immunoglobulin heavy chain variable region nucleotide sequence operably linked to an immunoglobulin heavy chain constant region nucleotide sequence. 
     
     
         115 . The nucleotide molecule of  claim 114 , wherein the heavy chain constant region gene sequence is selected from a C H 1, a hinge, a C H 2, a C H 3, and a combination thereof. 
     
     
         116 . The nucleotide molecule of  claim 114 , wherein the immunoglobulin heavy chain constant region nucleotide sequence is a human immunoglobulin heavy chain constant region nucleotide sequence. 
     
     
         117 . An immunoglobulin polypeptide comprising an amino acid sequence encoded by the nucleotide molecule of  claim 117 . 
     
     
         118 . A host cell comprising the nucleotide molecule of  claim 107 . 
     
     
         119 . The host cell of  claim 118 , further comprising a nucleic acid sequence that encodes a human light chain variable domain. 
     
     
         120 . The host cell of  claim 119 , wherein the nucleic acid sequence that encodes a human light chain variable domain is operably linked to an immunoglobulin light chain constant region gene sequence. 
     
     
         121 . The host cell of  claim 120 , wherein the immunoglobulin light chain constant region gene sequence is a human immunoglobulin light chain constant region gene sequence. 
     
     
         122 . The host cell of  claim 119 , further comprising
 (i) an immunoglobulin heavy chain variable domain encoded by the immunoglobulin heavy chain variable region nucleotide sequence that comprises a human germline heavy chain V H  gene segment and/or a human germline heavy chain J H  gene segment and   (ii) the immunoglobulin light chain variable domain.   
     
     
         123 . An antigen-binding protein expressed by the host cell of  claim 122 , wherein the antigen-binding protein comprises
 (i) the immunoglobulin heavy chain variable domain, and   (ii) the immunoglobulin light chain variable domain.