2. Academic Validation
  3. An efficient, non-viral arrayed CRISPR screening platform for iPSC-derived myeloid and microglia models

An efficient, non-viral arrayed CRISPR screening platform for iPSC-derived myeloid and microglia models

  • Stem Cell Reports. 2025 Mar 11;20(3):102420. doi: 10.1016/j.stemcr.2025.102420.
Sonja Meier 1,  Anne Sofie Gry Larsen 2,  Florian Wanke 1,  Nicolas Mercado 3,  Arianna Mei 4,  Livia Takacs 1,  Eva Suszanna Mracsko 1,  Ludovic Collin 1,  Martin Kampmann 5,  Filip Roudnicky 6,  Ravi Jagasia 1
Affiliations

Affiliations

  • 1 Pharma Research and Early Development, Neuroscience and Rare Diseases, F. Hoffmann-La Roche Ltd, 4070 Basel, Switzerland.
  • 2 Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT, England; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, England.
  • 3 Pharma Research and Early Development, Immunology and Inflammation, F. Hoffmann-La Roche Ltd, 4070 Basel, Switzerland.
  • 4 Pharma Research and Early Development, Therapeutic Modalities, F. Hoffmann-La Roche Ltd, 4070 Basel, Switzerland.
  • 5 Department of Biochemistry and Biophysics, Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA 94158, USA.
  • 6 Pharma Research and Early Development, Therapeutic Modalities, F. Hoffmann-La Roche Ltd, 4070 Basel, Switzerland. Electronic address: filip.roudnicky@roche.com.
PMID: 39983727 DOI: 10.1016/j.stemcr.2025.102420
Abstract

Here, we developed a CRISPR-Cas9 arrayed screen to investigate lipid handling pathways in human induced pluripotent stem cell (iPSC)-derived microglia. We established a robust method for the nucleofection of CRISPR-Cas9 ribonucleoprotein complexes into iPSC-derived myeloid cells, enabling genetic perturbations. Using this approach, we performed a targeted screen to identify key regulators of lipid droplet formation dependent on Apolipoprotein E (apoE). We identify the Mammalian Target of Rapamycin Complex 1 (mTORC1) signaling pathway as a critical modulator of lipid storage in both APOE3 and apoE knockout microglia. This study is a proof of concept underscoring the utility of CRISPR-Cas9 technology in elucidating the molecular pathways of lipid dysregulation associated with Alzheimer's disease and neuroinflammation.

Keywords

APOE; CRISPR-Cas9 gene editing; arrayed genetic screening; iPSC-derived microglia; lipid accumulation; lipid droplet screen; lipid metabolism; lipid regulation; lysosome; mTORC1.

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