Engineering monocyte/macrophage specific glucocerebrosidase expression in human hematopoietic stem cells using genome editing

Determine the feasibility of using CRISPR/Cas9 and AAV to genetically engineer human hematopoietic stem/progenitor cells (HSPCs) to express glucocerebrosidase under a macrophage-specific promoter as a curative therapy for Gaucher Disease.
Conclusion:
CRISPR/Cas9 and AAV achieve efficient targeted integration of human glucocerebrosdiase-expressing cassettes into human HSPCs. Targeted HSPCs exhibit mcarophage-specific cassette expression and are capable of long-term multi-lineage engraftment (including tissue-resident macrophages) in the NSG murine model . Negative controls (such as mock-treated cells) were used to draw marker-positive gates; cells treated with AAV vector alone (-RNP) were used to determine background episomal vector expression; compensation controls and FMOS were used where appropriate