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

被引：4

作者：

Chadarevian, Jean Paul ^{[1
,2
,3
]}

Davtyan, Hayk ^{[1
,2
,3
]}

Lombroso, Sonia I. ^{[4
,5
]}

Bennett, F. Chris ^{[4
,6
]}

Blurton-Jones, Mathew ^{[1
,2
,3
]}

机构：

[1] Univ Calif Irvine, Dept Neurobiol & Behav, Irvine, CA 92697 USA

[2] Univ Calif Irvine, Inst Memory Impairments & Neurol Disorders, Irvine, CA 92697 USA

[3] Univ Calif Irvine, Sue & Bill Gross Stem Cell Res Ctr, Irvine, CA 92697 USA

[4] Univ Penn, Perelman Sch Med, Dept Psychiat, Philadelphia, PA USA

[5] Univ Penn, Dept Syst Pharmacol & Translat Therapeut, Philadelphia, PA USA

[6] Childrens Hosp Philadelphia, Div Neurol, Philadelphia, PA USA

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 03期

关键词：

Cell Culture; Cell Differentiation; CRISPR; Immunology; Model Organisms; Neuroscience; Single Cell; Stem Cells;

D O I：

10.1016/j.xpro.2023.102490

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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

引用

页数：18

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