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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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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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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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CRISPR as a Diagnostic Tool.

Seohyun Kim1, Sangmin Ji1, Hye Ran Koh1

  • 1Department of Chemistry, Chung-Ang University, Seoul 06974, Korea.

Biomolecules
|August 27, 2021
PubMed
Summary
This summary is machine-generated.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas system offers precise gene targeting for diagnostics. Its Cas enzymes and guide RNA detect various targets, including genes, microRNAs, and proteins, enabling diverse diagnostic applications.

Keywords:
CRISPR-CasDNA methylationaptamerdiagnosisgene detectionmicroRNAsingle nucleotide polymorphism

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas system is a powerful tool for specific gene targeting.
  • CRISPR-Cas systems utilize Cas enzymes and guide RNA (gRNA) for DNA or RNA cleavage based on gRNA sequence specificity.

Purpose of the Study:

  • To review various CRISPR-based diagnostic methods.
  • To focus on the activity of CRISPR-Cas systems and target types in diagnostics.
  • To briefly introduce CRISPR-based diagnostics without target amplification.

Main Methods:

  • Review of CRISPR-Cas system activities including target-specific binding, cleavage, and trans-cleavage.
  • Analysis of CRISPR-Cas applications in detecting nucleic acids (genes, microRNAs, genetic variations) and non-nucleic acid targets (proteins).

Main Results:

  • CRISPR-Cas system's target-specific binding enables diverse diagnostic applications.
  • Cas12a and Cas13a exhibit trans-cleavage activity, enhancing diagnostic potential.
  • CRISPR-based methods can detect genetic variations like single nucleotide polymorphism and DNA methylation.

Conclusions:

  • The CRISPR-Cas system's versatility makes it a promising platform for developing advanced diagnostic tools.
  • CRISPR-based diagnostics can identify a wide range of disease-related targets with high specificity.
  • Further exploration of CRISPR-Cas activities and target forms will expand its diagnostic utility.