JP-2022534560-A5 -

Dates

Publication Date

20230530

Application Date

20200605

Description

In conclusion, hFIX was detected in the plasma of Alb +/hu /F9 -/- mice at 1, 3, and 4 weeks, and expressed hFIX-R338L was found to be functional because thrombin was generated in the TGA assay and aPTT coagulation time was improved. The present invention provides, for example, the following items: (Item 1) A non-human animal whose genome contains a humanized endogenous albumin locus, wherein a segment of the endogenous albumin locus has been deleted and replaced with a corresponding human albumin sequence. (Item 2) The non-human animal described in item 1, wherein the humanized endogenous albumin gene locus encodes a protein containing human serum albumin peptide. (Item 3) The non-human animal according to item 1 or 2, wherein the humanized endogenous albumin gene locus encodes a protein containing human albumin propeptide. (Item 4) A non-human animal as described in any one of the preceding items, wherein the humanized endogenous albumin gene locus encodes a protein containing human albumin signal peptide. (Item 5) A non-human animal according to any one of the preceding items, wherein the region of the endogenous albumin locus, which includes both coding and non-coding sequences, is deleted and replaced with a corresponding human albumin sequence, which also includes both coding and non-coding sequences. (Item 6) A non-human animal according to any one of the preceding items, wherein the humanized endogenous albumin locus includes the endogenous albumin promoter, and the human albumin sequence is operably linked to the endogenous albumin promoter. (Item 7) A non-human animal according to any one of the preceding items, wherein at least one intron and at least one exon of the endogenous albumin gene locus are deleted and replaced with the corresponding human albumin sequence. (Item 8) A non-human animal as described in any one of the preceding items, wherein the entire albumin coding sequence of the endogenous albumin locus is deleted and replaced with the corresponding human albumin sequence. (Item 9) The non-human animal described in item 8, wherein the region of the endogenous albumin locus from the start codon to the stop codon is deleted and replaced with the corresponding human albumin sequence. (Item 10) A non-human animal as described in any one of the preceding items, wherein the humanized endogenous albumin gene locus includes the human albumin 3' untranslated region. (Item 11) A non-human animal as described in any one of the preceding items, wherein the endogenous albumin 5' untranslated region is not deleted and is not replaced by the corresponding human albumin sequence. (Item 12) The region of the endogenous albumin locus from the start codon to the stop codon is deleted and replaced with a human albumin sequence including the corresponding human albumin sequence and the human albumin 3' untranslated region, and The endogenous albumin 5' untranslated region is not deleted, nor is it replaced by the corresponding human albumin sequence, and A non-human animal as described in any one of the preceding items, wherein the endogenous albumin promoter is not deleted and is not replaced by the corresponding human albumin sequence. (Item 13) (i) The human albumin sequence at the humanized endogenous albumin locus contains a sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence described in Sequence ID No. 35, or (ii) The humanized endogenous albumin locus codes for a protein that contains a sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% identical to the sequence described in Sequence ID No. 5, (iii) The humanized endogenous albumin locus contains a coding sequence which is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence described in Sequence ID No. 13, or (iv) The non-human animal according to any one of the preceding items, wherein the humanized endogenous albumin gene locus contains a sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence described in Sequence ID No. 17 or 18. (Item 14) A non-human animal as described in any one of the preceding items, wherein the humanized endogenous albumin locus does not contain a selection cassette or reporter gene. (Item 15) The non-human animal described in any one of the preceding items is homozygous for the humanized endogenous albumin locus. (Item 16) The non-human animal described in any one of the preceding items, wherein the non-human animal contains the humanized endogenous albumin gene locus in its germline. (Item 17) A non-human animal described in any one of the preceding items, wherein the aforementioned non-human animal is a mammal. (Item 18) The non-human animal described in item 17, wherein the non-human animal is a rat or a mouse. (Item 19) The no