CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model

Abstract

Summary: Chimeric mouse models have recently been developed to study human microglia in vivo. However, widespread engraftment of donor microglia within the adult brain has been challenging. Here, we present a protocol to introduce the G795A point mutation using CRISPR-Cas9 into the CSF1R locus of human pluripotent stem cells. We also describe an optimized microglial differentiation technique for transplantation into newborn or adult recipients. We then detail pharmacological paradigms to achieve widespread and near-complete engraftment of human microglia.For complete details on the use and execution of this protocol, please refer to Chadarevian et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Culture
• Cell Differentiation
• CRISPR
• Immunology
• Model Organisms
• Neuroscience