Gene editing in hematopoietic stem cells: tools for tracking, studying gene function, disease modeling, and potential promise for gene therapy
Hematopoietic stem cells (HSCs) are responsible for regeneration of all blood and immune cells over the lifespan through the process of gradual multilineage differentiation. Being the first adult stem cells discovered, the often serve as a paradigm for other stem cell systems. They are also the first stem cell type to be used clinically, and HSC transplantations are widely used for treating congenital blood diseases and to replenish blood production after cancer treatment.
CRISPR/Cas9 system represents a new fast and flexible way to investigate these cells and hematopoietic functioning overall in unprecedented details, allowing development of gene knockouts and knock-ins, gene tracking, as well as disease modeling in human HSCs where knockout models are not readily available. Further, it represents an attractive gene therapy option that would be based on transient transfection of patients cells that, unlike lentiviral systems, avoids stable integration of virus into the genome that can potentially can lead to insertional mutagenesis and leukemic transformation.
We developed a wide range of flexible tools and protocols that allow controlled gene editing in the hematopoietic system. We demonstrate efficient generation of knock-outs and knock-ins that can be used for analysis of gene function, and fluorescent labeling of gene of interest (that can be followed up in liquid cultures, colony forming assays and transplantation setting). In combination with our novel StemPro culture media for HSC maintenance, our approach can be potentially be beneficial for gene therapy protocols, allowing editing of engraftable HSCs.
Jason Potter graduated from Cornell and joined Life Technologies as a research scientist. Since then he has led development and evolution of enzymes including SuperScript III and other reverse transcriptase and polymerases at Invitrogen/Life Technologies. He has led groups focusing on synthetic genes and error correction. He now leads the Cell Biology Genome Editing R&D team in the Life Sciences Solutions Group of Thermo Fisher Scientific in Carlsbad, CA. His group is focused on developing tools for the entire genome editing workflow, specifically Gene Knock in and Knock out, Delivery, Analytics of Off Target events, and Cell Engineering.