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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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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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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
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Related Experiment Video

Updated: Apr 19, 2026

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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Modularized CRISPR/dCas9 effector toolkit for target-specific gene regulation.

Michael Agne1, Ilona Blank, Alica J Emhardt

  • 1iGEM Team Freiburg 2013, ‡BIOSS Centre for Biological Signalling Studies, University of Freiburg , Schänzlestrasse 18, 79104 Freiburg, Germany.

ACS Synthetic Biology
|December 20, 2014
PubMed
Summary
This summary is machine-generated.

We created a CRISPR/Cas9 toolkit for precise control of mammalian gene expression. This system allows customizable gene activation or repression for research and therapeutic applications.

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Area of Science:

  • Molecular Biology
  • Gene Editing
  • Synthetic Biology

Background:

  • Controlling mammalian gene expression synergistically is crucial for tissue engineering, stem cell reprogramming, and fundamental biological research.
  • Existing synthetic transcription factors offer limited customization and standardization.

Purpose of the Study:

  • To develop a standardized, customizable toolkit for precise gene regulation in mammalian cells.
  • To enable synergistic control of gene expression using an engineered CRISPR/Cas9 system.

Main Methods:

  • Engineered a CRISPR/Cas9 system utilizing the RNA-guided dCas9 protein.
  • Implemented dCas9 as a programmable transcriptional activator or repressor.
  • Developed a modular RNAimer plasmid for facile assembly of single or multiple CRISPR RNAs targeting endogenous loci.

Main Results:

  • Successfully created a standardized toolkit for customizable gene regulation.
  • Demonstrated the ability to target endogenous loci for gene activation or repression.
  • The toolkit facilitates the assembly of CRISPR RNAs for complex gene control.

Conclusions:

  • The developed CRISPR/Cas9 toolkit provides a powerful and flexible platform for mammalian gene regulation.
  • This system has broad applications in synthetic biology, regenerative medicine, and basic research.
  • The modular design enhances usability and scalability for diverse research needs.