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CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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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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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Related Experiment Video

Updated: Sep 5, 2025

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
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CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

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Programmable Transcriptional Modulation with a Structured RNA-Mediated CRISPR-dCas9 Complex.

Miao He1, Xiang Zhou1, Zhigang Li1

  • 1School of Chemistry and Materials Science, Department of Polymer Science and Engineering, iChEM (Collaborative Innovation Center of Chemistry for Energy Materials), University of Science and Technology of China, Hefei, Anhui 230026, China.

Journal of the American Chemical Society
|July 6, 2022
PubMed
Summary

Researchers developed a novel CRISPR-dCas9 system using structured RNA for programmable gene control. This system offers higher activation efficiency and enables targeted gene regulation in response to microRNAs, improving cellular control.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Synthetic Biology

Background:

  • Existing multi-module dCas9 (CRISPR-associated protein 9) systems offer controllable transcriptional manipulation.
  • A need exists for internal control modules within CRISPR-dCas9 systems for enhanced programmability.
  • Current systems like dCas9-VPR are widely used but may have limitations in efficiency and control.

Purpose of the Study:

  • To engineer a novel multi-module CRISPR-dCas9 system with a programmable RNA component for internal control.
  • To enhance gene activation efficiency compared to traditional CRISPR-dCas9 activators.
  • To develop a microRNA-responsive transcriptional regulation platform for endogenous gene activation and cell identification.

Main Methods:

  • Development of a multi-module CRISPR-dCas9 system incorporating a structured RNA as a programmable control element.
  • Introduction of a microRNA sensor to create a dCas9-based platform responsive to endogenous microRNAs.
  • Application of the platform for selective identification of HCT116 cells within a mixed cell population.

Main Results:

  • The novel structured RNA component demonstrated superior control over dCas9-based gene regulation.
  • Achieved higher transcriptional activation efficiency compared to the commonly used dCas9-VPR system.
  • Successfully generated a microRNA-responsive platform enabling controllable activation of endogenous genes and selective cell identification.

Conclusions:

  • The developed multi-module CRISPR-dCas9 system provides a flexible and efficient platform for programmable gene regulation.
  • The integration of a structured RNA component enhances control and activation efficiency.
  • This platform offers potential applications in synthetic biology, diagnostics, and targeted gene therapy.