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Related Experiment Video

Updated: Jun 28, 2025

Detection of MicroRNAs in Microglia by Real-time PCR in Normal CNS and During Neuroinflammation
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An integrated toolkit for human microglia functional genomics.

Imdadul Haq1,2,3, Jason C Ngo1,2,3, Nainika Roy1,2,3

  • 1Center for Translational and Computational Neuroimmunology, Columbia University Medical Center, New York, NY, USA.

Stem Cell Research & Therapy
|April 10, 2024
PubMed
Summary
This summary is machine-generated.

We developed a rapid, 20-day protocol to generate authentic human induced pluripotent stem cell-derived microglia (iMG) for Alzheimer's disease research. This toolkit enables functional genomics studies and investigation of genetic variants in neurodegenerative diseases.

Keywords:
CRISPRChromatin accessibility (ATAC-Seq)Functional genomicsMicrogliaNeurodegenerative diseasesProteomicsiPSC-derived microglia (iMG)

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Area of Science:

  • Neuroscience
  • Immunology
  • Genetics

Background:

  • Microglia are crucial brain immune cells involved in development and diseases like Alzheimer's disease (AD).
  • Human induced pluripotent stem cell-derived microglia (iMG) are valuable models, but current protocols are lengthy and lack comprehensive characterization.
  • Existing methods limit gene function studies using CRISPR-Cas9 in iMG.

Purpose of the Study:

  • To develop an optimized, rapid protocol for generating high-quality iMG.
  • To create a toolkit for functional genomics studies in iMG.
  • To facilitate the investigation of genetic variants in neurodegenerative diseases.

Main Methods:

  • A streamlined two-step, 20-day protocol for iPSC differentiation into iMG with normal karyotype.
  • Characterization using single-cell RNA sequencing, ATAC-Seq, proteomics, and functional assays.
  • Incorporation of a drug-dependent CRISPR-ON/OFF system for controlled gene expression and an online multi-omic data platform.

Main Results:

  • Generated iMG closely mirroring human primary microglia transcriptomically, proteomically, and epigenetically.
  • Demonstrated iMG functionality including Ca2+ transients, cytokine response, migration, and phagocytosis of AD-related substrates.
  • Established iMG as suitable models for studying AD-associated single nucleotide variants (SNVs) in regulatory regions.

Conclusions:

  • The developed protocol rapidly and efficiently produces authentic iMG.
  • The toolkit supports robust microglial functional genomics and SNV investigation.
  • This advancement promises progress in neurodegenerative disease drug discovery and therapy.