CRISPR 生成 CSF1R-G795A 人类小胶质细胞，用于在嵌合小鼠模型中进行强大的小胶质细胞替代。
CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model.
机构信息
Department 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.
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.
出版信息
STAR Protoc. 2023 Sep 15;4(3):102490. doi: 10.1016/j.xpro.2023.102490. Epub 2023 Jul 29.
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)..
嵌合鼠模型最近被开发用于研究体内的人类小胶质细胞。然而，在成年大脑中广泛植入供体小胶质细胞一直具有挑战性。在这里，我们提出了一种使用 CRISPR-Cas9 将 G795A 点突变引入人多能干细胞 CSF1R 基因座的方案。我们还描述了一种优化的小胶质细胞分化技术，用于移植到新生或成年受者体内。然后，我们详细介绍了实现广泛且近乎完全植入人类小胶质细胞的药理学范例。有关该方案使用和执行的完整详细信息，请参阅 Chadarevian 等人。（2023 年）。
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