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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

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Published on: June 2, 2018

Functional Enhancer Screening in Single Cells.

Joris van Arensbergen¹, Bas van Steensel²

¹Division of Gene Regulation, Netherlands Cancer Institute, 1066CX Amsterdam, the Netherlands.

|April 22, 2017

Summary

This summary is machine-generated.

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Researchers developed Mosaic-seq, a new technology combining CRISPR interference (CRISPRi) and single-cell RNA sequencing (scRNA-seq). This method allows for high-throughput analysis of enhancer roles in gene regulation.

Area of Science:

Molecular Biology
Genomics
Gene Regulation

Background:

Enhancers are crucial regulatory DNA elements controlling gene expression.
Understanding enhancer function is vital for deciphering complex biological processes.
Current methods for assessing enhancer contributions are often low-throughput or indirect.

Purpose of the Study:

To introduce and validate Mosaic-seq, a novel technology for high-throughput enhancer activity assessment.
To enable the simultaneous measurement of enhancer activity and gene expression at the single-cell level.
To provide a powerful tool for dissecting the regulatory logic of enhancers.

Main Methods:

Mosaic-seq integrates CRISPR interference (CRISPRi) for targeted gene silencing with single-cell RNA sequencing (scRNA-seq).

Keywords:

CRISPRi barcoding enhancer functional screen gene regulation promoter single-cell RNA sequencing

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Last Updated: Mar 3, 2026

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

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Published on: June 2, 2018

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Published on: March 31, 2022

A Rapid In Vivo Bioassay for Developmentally Active Enhancers

A Rapid In Vivo Bioassay for Developmentally Active Enhancers

CRISPRi libraries are used to perturb potential enhancers in individual cells.

scRNA-seq quantifies gene expression changes in response to enhancer perturbations.

Main Results:

Mosaic-seq successfully identified functional enhancers and their target genes.
The technology demonstrated high-throughput capacity for analyzing numerous enhancers simultaneously.
Single-cell resolution allowed for the detection of cell-to-cell variability in enhancer function.

Conclusions:

Mosaic-seq is a powerful and scalable technology for studying enhancer function.
This method significantly advances the ability to map enhancer-gene links and understand gene regulation.
Mosaic-seq opens new avenues for research in developmental biology, disease, and synthetic biology.