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Related Concept Videos

Genetic Screens02:46

Genetic Screens

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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
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Related Experiment Video

Updated: Oct 20, 2025

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

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Inducible CRISPRa screen identifies putative enhancers.

Zhongye Dai1, Rui Li1, Yuying Hou1

  • 1State Key Laboratory of Experimental Hematology, The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Tianjin Key Laboratory of Cellular Homeostasis and Human Diseases, Department of Cell Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China.

Journal of Genetics and Genomics = Yi Chuan Xue Bao
|September 17, 2021
PubMed
Summary

Researchers developed an inducible CRISPR activation (CRISPRa) system in mouse stem cells to identify gene enhancers. This system successfully pinpointed functional enhancers, including one linked to human height, demonstrating its utility for high-throughput enhancer screening.

Keywords:
CRISPR activationCis-regulatory elementsEnhancerScreen

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A Rapid In Vivo Bioassay for Developmentally Active Enhancers
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Area of Science:

  • Genomics
  • Developmental Biology
  • Gene Regulation

Background:

  • Enhancers are crucial cis-regulatory DNA elements controlling gene expression and cell differentiation.
  • Identifying enhancers in their native cellular environments remains a significant challenge in molecular biology.

Purpose of the Study:

  • To develop a novel inducible CRISPR activation (CRISPRa) system for efficient and precise enhancer identification in mouse embryonic stem cells (ESCs).
  • To utilize this system to screen for functional enhancers near the T (Brachyury) gene, a key regulator of mesodermal development.

Main Methods:

  • Generation of a doxycycline (Dox)-inducible CRISPRa system (iVPR ESC) in mouse ESCs, enabling tunable and reversible gene activation.
  • Tiled CRISPRa screening across a 105-kilobase genomic region surrounding the T gene using specific guide RNAs.
  • Chromatin analysis to identify CRISPRa-responsive elements with enhancer-like features.

Main Results:

  • The inducible CRISPRa system allowed for precise, dose-dependent, and reversible activation of target genes.
  • Several putative enhancers were identified, including one associated with human height genetic variants.
  • Deletion of the identified enhancer region in ESCs impaired T gene activation and osteogenic differentiation.

Conclusions:

  • The developed inducible CRISPRa ESC line offers a powerful and convenient platform for high-throughput functional screening of enhancers.
  • This system facilitates the discovery of novel regulatory elements and their roles in development and disease.