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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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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 double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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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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DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
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CRISPR-Cap: multiplexed double-stranded DNA enrichment based on the CRISPR system.

Jeewon Lee1, Hyeonseob Lim1, Hoon Jang1

  • 1Department of Chemistry, Yonsei University, Seoul 03722, Republic of Korea.

Nucleic Acids Research
|September 15, 2018
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CRISPR-Cap is a novel CRISPR-based DNA enrichment method that is fast, scalable, and accurate. This new technology offers improved target enrichment for genomic DNA analysis, gene copy number measurement, and rare allele detection.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Traditional DNA target enrichment methods like PCR, hybridization capture, and molecular inversion probes often suffer from long experiment times, low throughput, and suboptimal enrichment quality.
  • These limitations hinder efficient genomic DNA analysis and application development.

Purpose of the Study:

  • To develop a simple, scalable, and efficient CRISPR-based method for DNA target enrichment.
  • To overcome the drawbacks associated with existing target enrichment techniques.

Main Methods:

  • CRISPR-Cap, a novel CRISPR-based DNA target enrichment technique, was developed.
  • The method involves two short experimental procedures, completed within two hours, for enriching double-stranded DNA (dsDNA) target regions.

Main Results:

  • CRISPR-Cap achieved an average enrichment of 355.7-fold for 10 target genes from Escherichia coli genomic DNA, with a maximum on-target ratio of 81% and high uniformity.
  • The method successfully measured gene copy numbers and detected rare alleles at frequencies as low as 1%.
  • Coding sequence regions of 20 genes from the human genome were also enriched.

Conclusions:

  • CRISPR-Cap offers a rapid and scalable alternative to conventional target enrichment methods.
  • This technique broadens the application of CRISPR technology in the field of target enrichment for genomic analysis.