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Published April 2015 | public
Journal Article

Targeted Gene Theraphy in the Treatment of X-Linked Hyper-IgM Syndrome

Abstract

X-linked hyper-IgM syndrome results in absent IgG, IgA, IgE and normal/elevated IgM due to defects in the CD40 ligand gene. HSCT is the only curative modality, but it carries significant risks, suggesting the need for improved methods of treatment. TALENs or CRISPRs, combined with the effective delivery of a homologous donor sequence containing normal CD40L DNA, will allow for targeted integration and provide physiologic expression of the endogenous CD40L gene to provide long-term immune reconstitution. TALENs targeting the 5′ UTR of the CD40L gene lead to allelic disruption of up to 31%at the target locus inK562 cells. In order to evaluate the capacity for targeted integration of a cassette at the cut site, cells were electroporated with the TALEN pair and a donor molecule with a promoterless GFP reporter gene flanked by homology arms. Expression of the GFP reporter was evaluated in Jurkat cells, with up to 10 % detected by flow cytometry and increasing to 22 % upon PHA activation. In addition, CRISPRs targeting a patient-specific mutation in intron 3 achieved >50 % gene disruption in K562 cells. Co-electroporation with a donormodified to contain a unique restriction enzyme site demonstrated site-specific gene integration. Site-specific modification at CD40L is achievable and physiologic expression of the endogenous CD40L gene could provide a viable therapy for XHIM.

Additional Information

© 2015 Springer.

Additional details

Created:
August 20, 2023
Modified:
October 23, 2023