Poster #: 106
Optimization of Non-viral Knock-in in Hematopoietic Stem Cells
Mentors: Jenny Lee; Ke Li, PhD and Brian Shy, MD, PhD
CRISPR/Cas9 is a genome editing technology that enables targeted modification of DNA sequences in living cells, and CRISPR Knock-in is an application of CRISPR to induce a double-stranded break at specific target sites to perform gene insertion through Homology Directed Repair (HDR). We hypothesize that high editing efficiencies in CD34+ Hematopoietic Stem Cells (HSCs) can be achieved with non-viral knock-in methods at safe harbor site AAVS1 through optimization of electroporation buffer, pulse codes, as well as HDR template concentration. HSC editing through CRISPR-Cas9 can help scientists develop new therapy methods to treat conditions such as sickle cell disease, β-thalassemia, and many primary immunodeficiencies. Furthermore, autologous editing of a patient’s own HSCs minimizes risks of graft-versus-host-disease (GVHD), increasing the safety and efficacy of treatments.