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CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Yanxia Liu1, Chen Yu2, Timothy Patrick Daley3
1Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA.
Cell Stem Cell
|October 16, 2018
Summary
Scientists used CRISPR activation to discover new factors that specify neuronal fate. This approach identified key genes and factor pairs that can reprogram cells into neurons, advancing our understanding of cell-fate determination.
Area of Science:
- Developmental Biology
- Genetics
- Stem Cell Biology
Background:
- Specifying neuronal fate is crucial for understanding lineage development and cellular reprogramming.
- Systematically identifying regulatory factors and their interactions has been a significant technical challenge.
Purpose of the Study:
- To develop and apply a CRISPR activation (CRISPRa) approach for the systematic identification of neuronal fate regulators.
- To create a genetic interaction map for neuronal differentiation.
Main Methods:
- Utilized a pooled CRISPRa screen in embryonic stem cells to activate endogenous transcription factors and regulators.
- Performed systematic CRISPR-based activation of factor pairs to map genetic interactions.
- Confirmed individual and combinatorial hits for their ability to induce neuronal fate.
Main Results:
- Identified novel regulators of neuronal fate, including epigenetic factors like Ezh2.
- Generated a genetic interaction map detailing factors involved in neuronal differentiation.
- Demonstrated that several factor pairs can directly reprogram fibroblasts into neurons with transcriptional similarity to endogenous neurons.
Conclusions:
- The study presents an unbiased discovery approach for identifying genes driving cell-fate acquisition.
- The findings provide valuable insights into lineage specification and cellular reprogramming.
- The developed CRISPRa method offers a powerful tool for systematic gene discovery in cell-fate determination.
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