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Related Concept Videos

CRISPR01:59

CRISPR

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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

CRISPR and crRNAs

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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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Homologous Recombination02:31

Homologous Recombination

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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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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Related Experiment Video

Updated: May 9, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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Catching CRISPR-Cas9 in Action.

Yingjie Chen1, Yuanhao Li1, Penghai Li1

  • 1College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China.

Journal of Chemical Theory and Computation
|May 5, 2025
PubMed
Summary
This summary is machine-generated.

Understanding CRISPR-Cas9

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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Related Experiment Videos

Last Updated: May 9, 2025

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • CRISPR-Cas9 genome editing requires atomic-level understanding of its active conformation.
  • The full R-loop structure and dynamics of Cas9 are not fully characterized.

Purpose of the Study:

  • To investigate the structural dynamics and catalytic mechanisms of CRISPR-Cas9.
  • To provide insights for designing improved CRISPR-Cas9 systems.

Main Methods:

  • Integrated computational investigation using molecular dynamics simulations.
  • Quantum mechanics/molecular mechanics (QM/MM) simulations were employed.

Main Results:

  • Identified conformational heterogeneity in Cas9 and the nontarget DNA strand.
  • Revealed a conformational barrier for HNH nuclease domain activation.
  • Elucidated distinct cleavage pathways for nontarget and target strands.

Conclusions:

  • Cas9 conformational dynamics influence R-loop resolution and cleavage efficiency.
  • Strategic modulation of Cas9 domains can enhance genome editing.
  • Findings support rational design of next-generation CRISPR-Cas9 tools.