Journal
NEURON
Volume 104, Issue 2, Pages 239-+Publisher
CELL PRESS
DOI: 10.1016/j.neuron.2019.07.014
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Funding
- Intramural Research Program of the NIH/NINDS, an NIH Director's New Innovator Award [NIH/NIGMS DP2 GM119139]
- NIH/NIA [R01 AG062359, R56 AG057528]
- NINDS Tau Center Without Walls [NIH/NINDS U54 NS100717]
- Allen Distinguished Investigator Award
- Chan-Zuckerberg Biohub Investigator Award
- Tau Consortium Investigator Award (Rainwater Charitable Foundation)
- NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH [ZICDE000729] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [ZIANS003155] Funding Source: NIH RePORTER
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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.
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