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Related Concept Videos

Genetic Screens02:46

Genetic Screens

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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
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The Ras Gene02:38

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The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...
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Related Experiment Video

Updated: Apr 15, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
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Eprobe-mediated screening system for somatic mutations in the KRAS locus.

Jun Atsumi1, Takeshi Hanami2, Yasuaki Enokida1

  • 1Departments of Thoracic and Visceral Organ Surgery, Gunma University Graduate School of Medicine, Maebashi, Japan.

Oncology Reports
|April 1, 2015
PubMed
Summary

A new Eprobe system offers highly sensitive detection of Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations in colorectal cancer (CRC). This method surpasses conventional techniques, improving diagnostic capabilities for KRAS mutations.

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Author Spotlight: Advancing the Detection of Low-Frequency Mutations in Cancer Tissues
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Area of Science:

  • Molecular Biology
  • Oncology
  • Genetics

Background:

  • Activating Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations predict resistance to epidermal growth factor receptor (EGFR) therapy in colorectal cancer (CRC).
  • Current methods for detecting KRAS mutations lack sensitivity, feasibility, and cost-effectiveness.
  • A need exists for a highly sensitive and user-friendly KRAS mutation detection system for clinical applications.

Purpose of the Study:

  • To develop and validate a novel fluorescence Eprobe-based real-time PCR system for sensitive detection of KRAS gene mutations.
  • To assess the sensitivity and usability of the Eprobe system compared to conventional methods.

Main Methods:

  • Development of fluorescence Eprobes complementary to wild-type (WT) KRAS and common KRAS mutations (codons 12 and 13).
  • Utilizing Eprobe-PCR to detect low-level KRAS mutations by suppressing WT amplification and enriching mutant amplicons.
  • Comparative analysis of Eprobe-PCR with Sanger sequencing and next-generation sequencing (NGS) in 92 CRC patient samples.

Main Results:

  • The Eprobe system demonstrated high sensitivity in detecting KRAS mutations (0.05-0.1% plasmid DNA, 1% genomic DNA).
  • Eprobe-PCR identified KRAS mutations in CRC samples that were undetectable by Sanger sequencing.
  • The Eprobe system exhibited higher sensitivity than Sanger sequencing for KRAS mutation detection in patient samples.

Conclusions:

  • The novel Eprobe-PCR system provides a highly sensitive, user-friendly, and cost-effective method for detecting KRAS mutations.
  • This technology holds significant potential for improving diagnostic accuracy and guiding therapeutic decisions in colorectal cancer treatment.
  • The Eprobe approach is a valuable tool for clinical diagnostics, particularly for identifying low-level KRAS mutations.