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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 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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Related Experiment Video

Updated: Feb 5, 2026

Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti
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CRISPR-typing PCR (ctPCR), a new Cas9-based DNA detection method.

Qiao Wang1, Beibei Zhang1, Xinhui Xu1

  • 1State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.

Scientific Reports
|September 22, 2018
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Summary

A new CRISPR/Cas9-based method, CRISPR-typing PCR (ctPCR), enables rapid and sensitive DNA detection and typing. This technique accurately identifies specific DNA targets, such as human papillomavirus (HPV) subtypes, in clinical samples.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Accurate DNA detection and typing are crucial for diagnostics.
  • Existing methods may lack speed, specificity, or sensitivity.
  • CRISPR/Cas9 technology offers novel possibilities for molecular detection.

Purpose of the Study:

  • To develop and validate a novel CRISPR/Cas9-based method for DNA detection and typing.
  • To establish a sensitive, specific, rapid, and easy-to-use DNA analysis technique.
  • To demonstrate the method's utility in identifying specific viral DNA, such as HPV.

Main Methods:

  • Developed CRISPR-typing PCR (ctPCR), a three-step method involving universal PCR (PCR1), Cas9 cutting/adaptor ligation (CAT), and general-specific PCR (PCR2).
  • Utilized Cas9 nuclease and guide RNA for specific DNA targeting.
  • Verified the method by detecting HPV16 and HPV18 L1 genes in various HPV subtypes and clinical samples.

Main Results:

  • The ctPCR method demonstrated high sensitivity and specificity in detecting and typing target DNA.
  • Successfully identified HPV16 and HPV18 L1 genes in multiple high-risk HPV subtypes.
  • Validated the method's effectiveness in clinical samples, including cervical carcinoma cells.

Conclusions:

  • CRISPR-typing PCR (ctPCR) is a novel and effective method for DNA detection and genotyping.
  • The technique offers a significant advancement for identifying specific DNA sequences, like viral infections.
  • This CRISPR/Cas9-based approach provides a powerful tool for molecular diagnostics.