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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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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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Tracking CRISPR's Footprints.

Lin Lin1, Yonglun Luo2,3,4,5

  • 1Department of Biomedicine, Aarhus University, Aarhus, Denmark.

Methods in Molecular Biology (Clifton, N.J.)
|March 27, 2019
PubMed
Summary
This summary is machine-generated.

CRISPR gene editing relies on tracking DNA cleavage and binding footprints. New indel detection methods improve CRISPR-Cas9 tool activity and specificity for biomedical research.

Keywords:
CRISPRCas9DSBIndel frequencyIndelsOff-target

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Area of Science:

  • Biomedical research
  • Molecular biology
  • Gene editing technologies

Background:

  • Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) systems have transformed biomedical research.
  • Detecting and quantifying indels (insertions/deletions) is crucial for CRISPR gene editing.
  • Indels are DNA footprints resulting from CRISPR-Cas9 induced double-stranded breaks (DSBs) repair.

Purpose of the Study:

  • To review and discuss strategies for tracking CRISPR's DNA footprints.
  • To analyze the advantages and limitations of various indel tracking methods.
  • To highlight the role of these methods in enhancing CRISPR-Cas9 efficiency and specificity.

Main Methods:

  • Review of scientific literature on CRISPR footprint tracking.
  • Analysis of methods for detecting and quantifying DNA indels.
  • Discussion of CRISPR-Cas9 DNA-binding footprints without DSBs.

Main Results:

  • Indel tracking methods significantly contribute to improving CRISPR-Cas9 activity.
  • Understanding CRISPR's DNA-binding footprints is essential for precise gene editing.
  • Various strategies exist for tracking CRISPR-induced DNA modifications.

Conclusions:

  • Effective tracking of CRISPR footprints is vital for advancing gene editing technologies.
  • Continued development of indel detection methods will enhance CRISPR-Cas9 applications.
  • This review provides insights into current strategies for monitoring CRISPR-Cas9 activity and specificity.