Mentor: Andre Larochelle, M.D. Ph.D. |
Mentor: Christi Salisbury-Ruf, Ph.D. |
Lab: Laboratory of Regenerative Therapies for Inherited Blood Disorders

Genetic Engineering of Human Hematopoietic Stem Cells with CRISPR-Cas9: The Role of Innate Immunity

Genetic engineering of human hematopoietic stem and progenitor cells (HSPCs) is a promising therapy for individuals with inherited blood disorders. This approach uses CRISPR-Cas9 gene editing for precision genome targeting and a genetic template to insert a corrective gene or repair disease causing mutations.  Template DNA is commonly delivered by viruses bioengineered to contain the gene of interest. Cells harvested from patients can then be corrected with these tools in vitro followed by re-infusion into the same patient by autologous transplant. Our overall goal is to improve the efficiency of this therapy in human HSPCs (

Innate immune responses restrict HSPC genetic engineering and contribute to individual variability in clinical outcomes. The objective of this summer project is to understand the mechanisms underpinning the innate immune block to gene editing in HSPCs and explore approaches to overcome this inhibition. These translational studies could have a direct impact on the design of upcoming HSPC gene editing clinical trials and globally improve human health. 

This project involves an array of experimental laboratory tools including but not limited to:

  • Molecular biology techniques: proteomics, transcriptomics, gene editing of human HSPCs in vitro using CRISPR-Cas9 based approaches, bioengineering of viral vectors, RT-qPCR, Western blots, cloning, plasmid preparation, DNA extraction.
  • Cell biology techniques: culture of human HSPCs, viral vector production, transduction of human HSPCs, flow cytometry.
  • Animal work: mouse transplantation, dissection, tissue collection and processing. 
  • Bioinformatics tools: gene set enrichment analysis (GSEA) and other tools for the interpretation of high-throughput proteomics datasets.