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Decoding Neuronal Diversification by Multiplexed Single-cell RNA-Seq
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DOI：10.1016/j.stemcr.2021.02.006.png)
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摘要
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Cellular reprogramming is driven by a defined set of transcription factors; however, the regulatory logic that underlies cell-type specifi-cation and diversification remains elusive. Single-cell RNA-seq provides unprecedented coverage to measure dynamic molecular changes at the single-cell resolution. Here, we multiplex and ectopically express 20 pro-neuronal transcription factors in human dermal fibroblasts and demonstrate a widespread diversification of neurons based on cell morphology and canonical neuronal marker expressions. Single-cell RNA-seq analysis reveals diverse and distinct neuronal subtypes, including reprogramming processes that strongly correlate with the developing brain. Gene mapping of 20 exogenous pro-neuronal transcription factors further unveiled key determinants responsible for neuronal lineage specification and a regulatory logic dictating neuronal diversification, including glutamatergic and cholinergic neurons. The multiplex scRNA-seq approach is a robust and scalable approach to elucidate lineage and cellular specification across various biolog-ical systems.
Keyword:
HUMAN FIBROBLASTS
DIRECT CONVERSION
DOPAMINERGIC-NEURONS
MOUSE
PAX6
GENERATION
NEUROTRANSMITTER
REGULATORS
INDUCTION
SCREEN
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