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Related Concept Videos

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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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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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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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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Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice
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CRISPR/Cas9 in the Chicken Embryo.

Valérie Morin1, Nadège Véron2, Christophe Marcelle3,4

  • 1Institut NeuroMyoGène, INMG, Faculty of Medicine Laënnec, University Lyon1, Bâtiment B, 7 rue Guillaume Paradin, 69008, Lyon, France.

Methods in Molecular Biology (Clifton, N.J.)
|August 16, 2017
PubMed
Summary
This summary is machine-generated.

This study simplifies CRISPR/Cas9 genome editing combined with in vivo electroporation for effective gene loss-of-function in chicken embryos. This technique holds promise for treating genetic diseases.

Keywords:
CRISPRCas9Chicken embryoElectroporationgRNA

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Area of Science:

  • Biomedical Sciences
  • Genetics
  • Developmental Biology

Background:

  • Genome editing technologies, particularly CRISPR/Cas9, are revolutionizing biomedical research.
  • CRISPR/Cas9 enables targeted modification of genomes across diverse organisms.
  • Previous work demonstrated CRISPR/Cas9 combined with in vivo electroporation in chicken embryos.

Purpose of the Study:

  • To present a simplified and effective method for gene loss-of-function studies.
  • To optimize CRISPR/Cas9 and in vivo electroporation for somatic cell gene editing in developing embryos.

Main Methods:

  • Utilized the CRISPR/Cas9 system for targeted genome modification.
  • Employed in vivo electroporation for delivery into somatic cells of developing chicken embryos.
  • Developed a simplified protocol based on previously described techniques.

Main Results:

  • Achieved effective gene loss-of-function in chicken embryo somatic cells.
  • Demonstrated the utility of the simplified CRISPR/Cas9 and electroporation technique.
  • Validated the method for inhibiting gene function in a developmental context.

Conclusions:

  • The simplified CRISPR/Cas9 and in vivo electroporation technique is effective for gene function studies.
  • This method offers a promising approach for future therapeutic strategies targeting genetic diseases.
  • The technique is applicable to a wide spectrum of organisms and gene targets.