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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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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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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.
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Related Experiment Video

Updated: May 21, 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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Bacterial directed evolution of CRISPR base editors.

Reilly Q Mach1, Shannon M Miller1

  • 1The Scripps Research Institute.

Methods in Enzymology
|March 22, 2025
PubMed
Summary
This summary is machine-generated.

Directed evolution enhances CRISPR base editors for mammalian genome editing. This method uses bacterial evolution for efficient and flexible base editor engineering.

Keywords:
Base editingCRISPRDirected evolutionPrecision genome editingProtein engineering

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Related Experiment Videos

Last Updated: May 21, 2025

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genome Engineering

Background:

  • CRISPR-based genome editing, including base editing, has advanced therapeutic potential.
  • Many CRISPR enzymes require engineering for efficient mammalian genome editing.

Purpose of the Study:

  • To describe a method for bacterial directed evolution of CRISPR base editors.
  • To enhance the compatibility of base editors with mammalian cells.

Main Methods:

  • Plate-based discrete evolution was employed for enzyme engineering.
  • Bacterial systems were utilized for directed evolution of CRISPR base editors.

Main Results:

  • The described method offers a balance of ease of use and engineering power.
  • The bacterial directed evolution approach provides flexibility for base editor applications.

Conclusions:

  • Bacterial directed evolution is an effective strategy for engineering CRISPR base editors.
  • This method facilitates the development of more compatible and potent genome editing tools.