Composition of the Treatment of X-linked Chronic Granulomatous Disease (X-CGD) Deficiency in Human Hematopoietic Stem and Progenitor Cells (HSPCs)

Genome editing of human hematopoietic stem and progenitor cells (HSPCs) has the potential to create a new class of medication for the treatment of inherited and acquired genetic diseases of the blood and immune system. Researchers at Stanford have optimized a genome editing method in HSPCs using CRISPR/Cas9 in combination with AAV6-medicated homologous recombination (HR). The Cas9 nuclease and a short guide RNA (sgRNA) were delivered to the target genomic sequence. The double-stranded DNA break made by Cas9 was repaired by HR with a designed donor DNA template in the AAV6 vector that contains the desired genetic modification. Using this new method, researchers were able to achieve highly efficient editing, where both single nucleotide and several kilobases of DNA can be changed.
The researchers have applied the method to monogenic diseases, and have developed sgRNA and AAV6 donor DNA sequences that work well in HSPCs for several severe combined immunodeficiency disorders (SCID), including X-linked Chronic Granulomatous Disease (X-CGD) Deficiency.