US 12,376,573 B2
Genetically modified mice comprising humanized cellular immune system components with improved diversity of TCRB repertoire
Chunguang Guo, Briarcliff Manor, NY (US); Naxin Tu, Pleasantville, NY (US); and Andrew J. Murphy, Croton-on-Hudson, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Mar. 30, 2022, as Appl. No. 17/709,150.
Claims priority of provisional application 63/168,774, filed on Mar. 31, 2021.
Prior Publication US 2022/0322648 A1, Oct. 13, 2022
Int. Cl. A01K 67/027 (2024.01); A01K 67/0278 (2024.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C07K 14/73 (2006.01); C07K 14/74 (2006.01); C12N 5/0735 (2010.01)
CPC A01K 67/0278 (2013.01) [C07K 14/7051 (2013.01); C07K 14/70514 (2013.01); C07K 14/70517 (2013.01); C07K 14/70539 (2013.01); C12N 5/0606 (2013.01); A01K 2217/072 (2013.01); A01K 2227/105 (2013.01); A01K 2267/0387 (2013.01)] 23 Claims
 
1. A mouse cell comprising:
(A) an unrearranged T cell receptor (TCR) α variable region sequence that:
(i) comprises unrearranged human TCR α (TCRA) gene segments that comprise at least one functional unrearranged human T cell variable region Vα gene segment and at least one functional unrearranged human T cell variable region Jα gene segment, and
(ii) is operably linked to a mouse TCR α constant gene sequence, which is optionally at an endogenous mouse TCR α variable gene locus, and
(B) an unrearranged TCR β variable region sequence that:
(i) comprises unrearranged human TCR β (TCRB) gene segments that comprise at least one functional unrearranged human T cell variable region Vβ gene segment, a functional unrearranged human T cell variable region Dβ gene segment and a functional unrearranged human T cell variable region Jβ gene segment from each of a TCRBDJ1 cluster and a TCRBDJ2 cluster, and
(ii) is operably linked to a mouse TCR β constant gene sequence, which is optionally at an endogenous mouse TCR β variable gene locus,
wherein the unrearranged TCR β variable region sequence further comprises: (i) a mouse TCRBDJ1 non-coding sequence between the functional unrearranged human T cell variable region Dβ gene segment and the functional unrearranged human T cell variable region Jβ gene segment from the TCRBDJ1 cluster and (ii) a mouse TCRBDJ2 non-coding sequence between the functional unrearranged human T cell variable region Dβ gene segment and the functional unrearranged human T cell variable region Jβ gene segment from the TCRBDJ2 cluster,
wherein each of the unrearranged TCR α variable region sequence and the unrearranged TCR β variable region sequence is capable of rearranging in a T cell to encode a human or humanized T cell receptor variable α domain and a human or humanized T cell receptor variable β domain, respectively.