학술논문

CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model
Document Type
article
Source
STAR Protocols. 4(3)
Subject
Medical Biotechnology
Biomedical and Clinical Sciences
Stem Cell Research
Stem Cell Research - Embryonic - Human
Transplantation
Stem Cell Research - Induced Pluripotent Stem Cell
Neurosciences
5.2 Cellular and gene therapies
Adult
Animals
Mice
Infant
Newborn
Humans
Microglia
Brain
Disease Models
Animal
Pluripotent Stem Cells
Point Mutation
CRISPR
Cell Culture
Cell Differentiation
Immunology
Model Organisms
Neuroscience
Single Cell
Stem Cells
Language
Abstract
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.