CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model
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Updated: Jul 20, 2025
Transplantation of Human Induced Pluripotent Stem Cell-Derived Microglia in Immunocompetent Mice Brain via Non-Invasive Transnasal Route
Published on: May 31, 2022
Jean Paul Chadarevian1, Hayk Davtyan2, Sonia I Lombroso3
1Department of Neurobiology & Behavior, University of California Irvine, Irvine, CA 92697, USA; Institute for Memory Impairments and Neurological Disorders, University of California Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, CA 92697, USA.
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.
Insights
Researchers developed a new protocol for widespread human microglia engraftment in mouse models. This method uses CRISPR-Cas9 gene editing and optimized cell differentiation for improved in vivo study of human microglia.
Area of Science:
- Neuroscience
- Immunology
- Genetics
Background:
- Chimeric mouse models are crucial for studying human microglia in vivo.
- Achieving widespread engraftment of donor microglia in adult mouse brains remains a significant challenge.
Purpose of the Study:
- To present a protocol for enhanced human microglia engraftment in mouse models.
- To overcome limitations in studying human microglia function in vivo.
Main Methods:
- Utilized CRISPR-Cas9 to introduce a G795A point mutation into the CSF1R locus of human pluripotent stem cells.
- Developed an optimized microglial differentiation technique for transplantation.
- Employed pharmacological strategies to promote widespread microglia engraftment.
Main Results:
- Successfully introduced the G795A point mutation in human pluripotent stem cells.
- Demonstrated an optimized protocol for microglial differentiation and transplantation.
- Achieved widespread and near-complete engraftment of human microglia in mouse recipients.
Conclusions:
- The presented protocol facilitates robust human microglia engraftment in mouse models.
- This advancement enables more effective in vivo research on human microglia.
- The findings contribute to understanding human microglia biology and potential therapeutic targets.
