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Related Concept Videos

CRISPR01:59

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Related Experiment Video

Updated: Jan 26, 2026

Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR
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Gene activation by a CRISPR-assisted trans enhancer.

Xinhui Xu1, Jinliang Gao1, Wei Dai1

  • 1State Key Laboratory of Bioelectronics, Southeast University, Nanjing, China.

Elife
|April 12, 2019
PubMed
Summary

This study introduces a novel CRISPR-assisted trans enhancer strategy to significantly boost gene activation efficiency. This new method overcomes limitations of current dCas9 activators, offering higher performance in various cell types.

Keywords:
CMV enhancerCRISPR/dCas9biochemistrycell biologychemical biologygene activationhumantrans enhancer

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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biotechnology

Background:

  • Deactivated CRISPR/Cas9 (dCas9) is a key tool for gene activation.
  • Existing dCas9-based activators face limitations in achieving satisfactory activity levels.

Purpose of the Study:

  • To develop a novel CRISPR-assisted trans enhancer strategy for highly efficient gene expression activation.
  • To overcome the limitations of current dCas9 activators.

Main Methods:

  • Combined dCas9-VP64/sgRNA with the CMV enhancer.
  • Developed two systems for recruiting the CMV enhancer in trans: dCas9-VP64/csgRNA-sCMV and dCas9-VP64-GAL4/sgRNA-UAS-CMV.
  • Utilized oligonucleotide annealing and GAL4-UAS binding for enhancer recruitment.

Main Results:

  • The trans enhancer mimicked natural cis enhancers in activating gene transcription.
  • Demonstrated high activation efficiency for both exogenous reporter genes and endogenous genes.
  • Achieved significantly higher activation efficiency compared to existing dCas9 activators across various cell types.

Conclusions:

  • The CRISPR-assisted trans enhancer strategy offers a powerful new approach for high-efficiency gene activation.
  • This method provides a significant improvement over current dCas9-based gene activators.
  • The strategy is effective in diverse cellular contexts for both reporter and endogenous gene targets.