Granted Patent
B2
US 12,201,096 · App. 18/347,009 · Granted Jan 21, 2025
Non-human animals comprising a humanized coagulation factor 12 locus
Inventors:
Yajun Tang (White Plains, NY); Dan Chalothorn (New York, NY); Lyndon Mitnaul (Piscataway, NJ); Lori Morton (Chappaqua, NY); Daria Zamolodchikov (Hastings-on-Hudson, NY); Nicole Alessandri-Haber (Rye Brook, NY); Lynn Macdonald (Harrison, NY)
Assignee:
REGENERON PHARMACEUTICALS, INC.
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Abstract
Non-human animal genomes, non-human animal cells, and non-human animals comprising a humanized coagulation factor XII (F12) locus and methods of making and using such non-human animal genomes, non-human animal cells, and non-human animals are provided. Non-human animal cells or non-human animals comprising a humanized F12 locus express a human coagulation factor XII protein or a chimeric coagulation factor XII protein, fragments of which are from human coagulation factor XII. Methods are provided for using such non-human animals comprising a humanized F12 locus to assess in vivo efficacy of human-coagulation-factor-XII-targeting reagents such as nuclease agents designed to target human F12. A short isoform of F12 that is produced locally in the brain, and methods of using the short isoform, are also provide.
Claims (19)
1. A method of assessing expression of a short F12 mRNA isoform in a biological sample, comprising:
(a) detecting and quantifying RNA transcripts using primers and/or probes against one or more of exons 9-14 of an F12 gene; and
(b) detecting and quantifying RNA transcripts using primers and/or probes against one or more of exons 1-6 of the F12 gene
wherein the short isoform is detected by primers and/or probes against exons 9-14 but not by primers and/or probes against exons 1-6.
2. The method of claim 1 , wherein the biological sample is from a brain.
3. The method of claim 1 , wherein the biological sample comprises neurons.
4. The method of claim 1 , wherein the biological sample is from a human.
5. The method of claim 1 , wherein the biological sample is from a non-human animal with a humanized F12 locus.
6. The method of claim 1 , wherein the biological sample comprises human neurons.
7. The method of claim 1 , wherein the biological sample comprises neurons from a non-human animal with a humanized F12 locus.
8. The method of claim 1 , wherein step (a) comprises detecting and quantifying RNA transcripts using primers and/or probes against one or more of exons 11-14 of an F12 gene, wherein the short isoform is detected by primers and/or probes against exons 11-14 but not by primers and/or probes against exons 1-6.
9. The method of claim 8 , wherein the biological sample comprises human neurons.
10. The method of claim 8 , wherein the biological sample comprises neurons from a non-human animal with a humanized F12 locus.
11. The method of claim 1 , wherein the detecting and quantifying in steps (a) and (b) comprises quantitative PCR.
12. The method of claim 1 , further comprising assessing activity of the short F12 mRNA isoform.
13. The method of claim 12 , wherein assessing activity of the short F12 mRNA isoform comprises measuring hepatocyte growth factor (HGF)-tyrosine-protein kinase Met (Met) signaling activity.
14. The method of claim 12 , wherein assessing activity of the short F12 mRNA isoform comprises assessing activation of prekallikrein to kallikrein, assessing activation of FXI, assessing activation of pro-HGF to HGF, or assessing activation of the classical complement pathway.
15. The method of claim 14 , wherein the biological sample comprises human neurons.
16. The method of claim 14 , wherein the biological sample comprises neurons from a non-human animal with a humanized F12 locus.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Nov 4, 2024
From: TANG, YAJUN; CHALOTHORN, DAN; MITNAUL, LYNDON; MORTON, LORI; ZAMOLODCHIKOV, DARIA; ALESSANDRI-HABER, NICOLE; MACDONALD, LYNN
To: REGENERON PHARMACEUTICALS, INC.
Reel/Frame 069123/0201 →
Continuity (3)
Provisional Application
62829321
· Apr 4, 2019
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