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

• DOI: 10.1016/j.stemcr.2025.102420
• Publication Date: 2025-02-20T15:33:17Z
• Authors: Sonja Meier; Anne Sofie Gry La...; Florian Wanke; Nicolas Mercado; Arianna Mei; Livia Takacs; Eva Suszanna Mracsko; Ludovic Collin; Martin Kampmann; Filip Roudnicky; Ravi Jagasia

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