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Related Concept Videos

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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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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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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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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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Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
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CRISPR-Cas12a trans-cleaves DNA G-quadruplexes.

Ying Li1, Tao Li1, Bi-Feng Liu2

  • 1State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan National Laboratory for Optoelectronics, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences - Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, 430071, China. liying@wipm.ac.cn ml.liu@wipm.ac.cn and University of Chinese Academy of Sciences, Beijing, 10049, China.

Chemical Communications (Cambridge, England)
|September 23, 2020
PubMed
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The CRISPR-Cas12a system can now cleave DNA G-quadruplexes (G4). This discovery enhances CRISPR-Cas12a and G4 applications in biosensing and biochemistry.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • DNA G-quadruplexes (G4) are non-canonical DNA structures implicated in various biological processes.
  • CRISPR-Cas12a is a versatile gene-editing tool with nuclease activity.

Purpose of the Study:

  • To investigate the potential of the CRISPR-Cas12a system to cleave DNA G-quadruplexes (G4).

Main Methods:

  • Utilized FRET, Circular Dichroism (CD), gel electrophoresis, and Nuclear Magnetic Resonance (NMR) to analyze cleavage activity.
  • Tested cleavage on human telomere G4 and TBA G4 structures.

Main Results:

  • Demonstrated for the first time that the activated CRISPR-Cas12a system exhibits trans-cleavage activity on DNA G-quadruplexes.
  • Verified the cleavage activity using multiple biophysical and biochemical techniques.

Conclusions:

  • The CRISPR-Cas12a system's ability to cleave G4 structures opens new possibilities.
  • This finding advances the application of CRISPR-Cas12a and G4 in biosensing and biochemistry.