CRISPR-Cas9 gene editing of human hematopoietic stem and progenitor (HSPC) cells holds great promise for disease treatment by cellular therapy and other methodologies. However, the variables that can affect treatment of relevant diseases in the real world are still being elucidated. A critical part of this involves implementing an optimized gene editing and cell culture methodology. In this webinar, Dr. Marta Walasek and Ms. Danielle Nguyen Truong describe tips and tricks for effective genome editing of CD34⁺ cells, methods to evaluate genome editing efficiency, as well as optimal pre- and post- editing culture conditions to maintain HSPC function and long-term editing effects.