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Establishment of single-cell screening system for the rapid identification of transcriptional modulators involved in direct cell reprogramming.

ABSTRACT: Combinatorial interactions of transcription modulators are critical to regulate cell-specific expression and to drive direct cell reprogramming (e.g. trans-differentiation). However, the identification of key transcription modulators from myriad of candidate genes is laborious and time consuming. To rapidly identify key regulatory factors involved in direct cell reprogramming, we established a multiplex single-cell screening system using a fibroblast-to-monocyte transition model. The system implements a single-cell 'shotgun-transduction' strategy followed by nested-single-cell-polymerase chain reaction (Nesc-PCR) gene expression analysis. To demonstrate this, we simultaneously transduced 18 monocyte-enriched transcription modulators in fibroblasts followed by selection of single cells expressing monocyte-specific CD14 and HLA-DR cell-surface markers from a heterogeneous population. Highly multiplex Nesc-PCR expression analysis revealed a variety of gene combinations with a significant enrichment of SPI1 (86/86) and a novel transcriptional modulator, HCLS1 (76/86), in the CD14(+)/HLA-DR(+) single cells. We could further demonstrate the synergistic role of HCLS1 in regulating monocyte-specific gene expressions and phagocytosis in dermal fibroblasts in the presence of SPI1. This study establishes a platform for a multiplex single-cell screening of combinatorial transcription modulators to drive any direct cell reprogramming.

SUBMITTER: Shin JW

PROVIDER: S-EPMC3505982 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Cardiac-mimetic cell-culture system for direct cardiac reprogramming.

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| S-EPMC6815967 | biostudies-literature

A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types.

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Rapid Chromatin Switch in the Direct Reprogramming of Fibroblasts to Neurons.

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| S-EPMC5646379 | biostudies-literature

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| S-EPMC8161510 | biostudies-literature

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