IP Library › Granted Patent US 12,735,720
Granted Patent B2
US 12,735,720 · App. 17/059,191 · Granted Sep 15, 2026

Gene editing of monogenic disorders in human hematopoietic stem cells—correction of x-linked hyper-IGM syndrome (XHIM)

Inventors: Joseph Long (Studio City, CA); Donald B. Kohn (Tarzana, CA); Caroline Y. Kuo (Beverly Hills, CA)
Assignee: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
C12N15/86A61K38/1793A61K40/11A61K40/40A61K48/0066A61P37/04C12N5/0636C12N9/22C12N15/11C12N15/907C12N2310/14C12N2310/20C12N2740/15043C12N2750/14143C12N2800/80
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Quick Facts
Patent No.
US 12,735,720
App. No.
17/059,191
Granted
Sep 15, 2026
Kind
B2
Abstract

Described herein are methods of treating X-Linked Hyper-IgM Syndrome (XHIM) in a mammal. Methods include: (i) providing differentiated T cells and/or stem/progenitor cells obtained from the mammal; and (ii) performing a targeted insertion of a corrective CD40L cDNA at the CD40LG gene locus in the differentiated T cells and/or stem/progenitor cells. The targeted insertion provides a corrected CD40LG gene and places the corrective CD40L cDNA downstream and operably linked to the endogenous CD40LG enhancer/promoter. The method further includes (iii) introducing these recombinant cells into the mammal, wherein the corrected CD40LG gene is expressed in a physiologically regulated manner.

Claims (12)

1 . A method of engraftment of edited T cells and/or stem/progenitor cells in a mammal, said method comprising:

providing differentiated T cells and/or stem/progenitor cells from said mammal;

performing a targeted insertion of a corrective CD40L cDNA donor at the CD40LG gene locus in said cells, downstream and operably linked to the endogenous CD40LG enhancer/promoter, to provide regulated control of the corrective CD40L cDNA donor, wherein said corrective CD40L cDNA donor comprises an optimized CD40L cDNA sequence and the 3′ UTR of the CD40L gene, said donor comprising SEQ ID NO: 13, SEQ ID NO: 9, or SEQ ID NO: 11, and wherein said targeted insertion comprises transducing said cells with an AAV vector or a lentiviral vector comprising the corrective CD40L cDNA donor; and transfecting said cells in vitro, with

a nucleic acid encoding a guide RNA (gRNA) and CRISPR/Cas endonuclease where said gRNA targets the CD40L 5′ UTR; or

a ribonucleoprotein (RNP) complex comprising a guide RNA complexed to a CRISPR/Cas endonuclease, where said gRNA targets the CD40L 5′ UTR, and

wherein said gRNA comprises the sequence set forth in SEQ ID NO: 12; and

introducing said cells into said mammal;

wherein said method provides a corrected CD40LG gene expressed in a physiologically regulated manner.

2 . The method of claim 1 , wherein said corrective CD40L cDNA donor is flanked by homology arms.

3 . The method of claim 1 , wherein said corrective CD40L cDNA donor sequence is followed by the bGH polyA signal.

4 . The method of claim 3 , wherein said corrective CD40L cDNA donor sequence is flanked on its 5′ end by a 5′ homology arm that begins at the CRISPR/Cas cut site and extends at least 100 bp upstream; and/or following the bGH polyA signal it is flanked on its 3′ end by a 3′ homology arm that extends at least 200 bp, or at least 300 bp, or at least 350 bp, or at least 400 bp downstream of the CRISPR/Cas cut site.

5 . The method of claim 1 , wherein said cells comprise hematopoietic stem and progenitor cells (HSPC).

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 18, 2023
From: KOHN, DONALD B.; KUO, CAROLINE Y.; LONG, JOSEPH
To: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Reel/Frame 063359/0538 →
Continuity (3)
Provisional Application 62760448 · Nov 13, 2018
Provisional Application 62678218 · May 30, 2018
Related Publication 20230158110A1 · May 25, 2023
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