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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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CRISPR01:59

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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

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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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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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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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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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CRISPR Approaches to Small Molecule Target Identification.

Marco Jost1,2, Jonathan S Weissman1

  • 1Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, Center for RNA Systems Biology, University of California, San Francisco , San Francisco, California 94158, United States.

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|December 21, 2017
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Identifying how small molecules work is crucial for drug development. Chemical-genetic methods, especially those using CRISPR technology, systematically profile gene effects on drug sensitivity in mammalian systems.

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Area of Science:

  • Pharmacology
  • Genetics
  • Drug Discovery

Background:

  • Identifying the mechanisms of action for potential therapeutics is a significant hurdle in drug development.
  • Various methods exist, each with unique advantages and disadvantages.

Purpose of the Study:

  • To review methods for identifying small molecule mechanisms of action.
  • To focus on chemical-genetic approaches utilizing genetic perturbations to assess drug sensitivity.

Main Methods:

  • Review of existing methodologies for mechanism of action studies.
  • Emphasis on chemical-genetic profiling.
  • Application of CRISPR-based technologies for gene repression, induction, or deletion in mammalian systems.

Main Results:

  • CRISPR-based approaches have advanced the application of chemical-genetic methods in mammalian systems.
  • These methods allow for systematic profiling of genetic perturbation effects on drug sensitivity.

Conclusions:

  • Chemical-genetic methods, particularly with CRISPR, offer powerful tools for elucidating small molecule mechanisms of action.
  • Advancements in genetic perturbation technologies facilitate drug discovery and development research.