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Related Concept Videos

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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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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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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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.
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Related Experiment Video

Updated: Apr 15, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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Small molecule-triggered Cas9 protein with improved genome-editing specificity.

Kevin M Davis1, Vikram Pattanayak2, David B Thompson1

  • 11] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.

Nature Chemical Biology
|April 8, 2015
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Summary

Researchers developed a new Cas9 gene-editing tool controlled by a small molecule. This conditionally active Cas9 significantly enhances editing specificity, reducing off-target modifications in human cells.

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Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Biochemistry

Background:

  • Genome editing technologies like CRISPR-Cas9 offer powerful tools for genetic manipulation.
  • Enhancing the specificity of genome editing is crucial to minimize unintended alterations and improve therapeutic applications.
  • Controlling Cas9 activity post-delivery can increase precision at the target locus.

Purpose of the Study:

  • To develop a novel Cas9 nuclease system with enhanced specificity.
  • To create a Cas9 variant that can be conditionally activated by a small molecule.
  • To evaluate the specificity of the developed Cas9 system in human cells.

Main Methods:

  • Engineered Cas9 nucleases by inserting an evolved 4-hydroxytamoxifen-responsive intein.
  • Utilized a cell-permeable small molecule for conditional activation of Cas9.
  • Assessed genome modification specificity in human cells compared to wild-type Cas9.

Main Results:

  • Successfully developed conditionally active Cas9 nucleases activated by a small molecule.
  • Demonstrated that the engineered Cas9 systems exhibit significantly higher specificity.
  • Achieved up to 25-fold increased specificity in modifying target genomic sites in human cells compared to wild-type Cas9.

Conclusions:

  • Conditionally active Cas9 nucleases activated by small molecules represent a promising strategy for improving genome editing specificity.
  • This approach offers enhanced control over Cas9 activity, leading to more precise genome modification.
  • The developed system has the potential to reduce off-target effects in gene editing applications.