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

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.