Doctor of Medicine, Hanover School of Medicine (2011)
Staatsexamen, Julius Maximilians Univsitat (2010)
Matthew Porteus, Postdoctoral Faculty Sponsor
Genome Editing Using CRISPR/Cas9 and rAAV6 to Functionally Correct Wiskott-Aldrich Syndrome in Human HSPCs
CELL PRESS. 2018: 376–77
View details for Web of Science ID 000435342205038
Genome Editing of Long-Term Human Hematopoietic Stem Cells for X-Linked Severe Combined Immunodeficiency
SPRINGER/PLENUM PUBLISHERS. 2018: 365–66
View details for Web of Science ID 000431311600087
A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells.
2018; 24 (8): 1216–24
Translation of the CRISPR-Cas9 system to human therapeutics holds high promise. However, specificity remains a concern especially when modifying stem cell populations. We show that existing rationally engineered Cas9 high-fidelity variants have reduced on-target activity when using the therapeutically relevant ribonucleoprotein (RNP) delivery method. Therefore, we devised an unbiased bacterial screen to isolate variants that retain activity in the RNP format. Introduction of a single point mutation, p.R691A, in Cas9 (high-fidelity (HiFi) Cas9) retained the high on-target activity of Cas9 while reducing off-target editing. HiFi Cas9 induces robust AAV6-mediated gene targeting at five therapeutically relevant loci (HBB, IL2RG, CCR5, HEXB, and TRAC) in human CD34+ hematopoietic stem and progenitor cells (HSPCs) as well as primary T cells. We also show that HiFi Cas9 mediates high-level correction of the sickle cell disease (SCD)-causing p.E6V mutation in HSPCs derived from patients with SCD. We anticipate that HiFi Cas9 will have wide utility for both basic science and therapeutic genome-editing applications.
View details for DOI 10.1038/s41591-018-0137-0
View details for PubMedID 30082871