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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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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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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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CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis
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CRISPR as a strong gene editing tool.

Shengfu Shen1, Tiing Jen Loh2, Hongling Shen2

  • 1Willston Northampton School, Easthampton, MA 01027, USA.

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Summary
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CRISPR gene editing uses guide RNA and Cas9 to precisely cut DNA, enabling applications in agriculture and human genetic disease treatment. DNA repair mechanisms like NHEJ and HDR facilitate the editing process.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Clustered regularly-interspaced short palindromic repeats (CRISPR) technology has emerged as a powerful genetic editing tool.
  • Initially discovered in bacteria as a defense mechanism against viral infections.
  • CRISPR systems leverage RNA-guided nucleases for targeted DNA manipulation.

Purpose of the Study:

  • To elucidate the mechanism of CRISPR-Cas9 mediated gene editing.
  • To describe the roles of guide RNA, Cas9, and RNase III in the process.
  • To outline the DNA repair pathways involved in CRISPR-induced modifications.

Main Methods:

  • Identification of viral DNA by guide RNA (crRNA and tracrRNA).
  • Processing of pre-crRNA into crRNA by RNase III.
  • Formation of a crRNA:tracrRNA:Cas9 complex to guide DNA cleavage.
  • Analysis of DNA repair mechanisms: Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR).

Main Results:

  • CRISPR-Cas9 system facilitates targeted DNA cleavage.
  • NHEJ offers a simple, albeit random, DNA repair pathway.
  • HDR provides a more complex and accurate DNA repair mechanism.
  • The CRISPR-Cas9 system is versatile for various biological applications.

Conclusions:

  • CRISPR-Cas9 is an effective gene editing technology with significant potential.
  • Applications span diverse fields, including agriculture and human therapeutics for genetic diseases.
  • Understanding the underlying mechanisms and repair pathways is crucial for optimizing CRISPR technology.