Details

Autor(en) / Beteiligte

• Tian, Ruilin
• Gachechiladze, Mariam A.
• Ludwig, Connor H.
• Laurie, Matthew T.
• Hong, Jason Y.
• Nathaniel, Diane
• Prabhu, Anika V.
• Fernandopulle, Michael S.
• Patel, Rajan
• Abshari, Mehrnoosh
• Ward, Michael E.
• Kampmann, Martin

Titel

CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons

Ist Teil von

• Neuron (Cambridge, Mass.), 2019-10, Vol.104 (2), p.239-255.e12

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2019

Quelle

ScienceDirect

Beschreibungen/Notizen

• CRISPR/Cas9-based functional genomics have transformed our ability to elucidate mammalian cell biology. However, most previous CRISPR-based screens were conducted in cancer cell lines rather than healthy, differentiated cells. Here, we describe a CRISPR interference (CRISPRi)-based platform for genetic screens in human neurons derived from induced pluripotent stem cells (iPSCs). We demonstrate robust and durable knockdown of endogenous genes in such neurons and present results from three complementary genetic screens. First, a survival-based screen revealed neuron-specific essential genes and genes that improved neuronal survival upon knockdown. Second, a screen with a single-cell transcriptomic readout uncovered several examples of genes whose knockdown had strikingly cell-type-specific consequences. Third, a longitudinal imaging screen detected distinct consequences of gene knockdown on neuronal morphology. Our results highlight the power of unbiased genetic screens in iPSC-derived differentiated cell types and provide a platform for systematic interrogation of normal and disease states of neurons. [Display omitted] [Display omitted] •A CRISPR interference platform for genetic screens in human iPSC-derived neurons•Survival screens uncover genes essential for neurons, but not iPSCs or cancer cells•Single-cell RNA-seq screens reveal distinct neuronal roles for ubiquitous genes•Arrayed high-content screens uncover genes controlling neuronal morphology Tian, Gachechiladze, and Ludwig et al. present a CRISPR interference-based platform for genetic screens in human iPSC-derived neurons. This platform enables systematic elucidation of gene function in human neurons and reveals neuron-specific roles of genes for survival, transcriptomics states, and morphology.