2. Academic Validation
  3. CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model

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

  • STAR Protoc. 2023 Jul 29;4(3):102490. doi: 10.1016/j.xpro.2023.102490.
Jean Paul Chadarevian 1,  Hayk Davtyan 2,  Sonia I Lombroso 3,  F Chris Bennett 4,  Mathew Blurton-Jones 5
Affiliations

Affiliations

  • 1 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. Electronic address: jchadare@uci.edu.
  • 2 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.
  • 3 Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania, Philadelphia, PA, USA.
  • 4 Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Division of Neurology, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
  • 5 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. Electronic address: mblurton@uci.edu.
PMID: 37516973 DOI: 10.1016/j.xpro.2023.102490
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.

Keywords

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

Figures
Products
嗨！很高兴为您提供帮助！

尊敬的 MCE 客户您好，
请您选择所在区域，我们将转接对应客服为您服务！

热门区域

A

B

C

F

G

H

J

L

N

Q

S

T

X

Y

Z

热 A B C F G H J L N Q S T X Y Z