Korea’s first MRD diagnostic kit for Sanger Sequencing

CRISmono™ is a high-performance Sanger Sequencing genetic mutation detection kit developed for Minimal Residual Disease (MRD) examination and diagnostic monitoring.

Ultra-precision CRISPR (Clustered Regular Interspaced Short Palindromic Repeat) genetic scissors technology is applied. This technology amplifies mutant genes from slight amounts of cell-free DNA in blood and improves the accuracy of mutation detection.

Minimal Residual Disease (MRD) diagnosis

Cancer can cause recurrence if some cancer cells remain after treatment.

MRD diagnosis is essential in predicting the possibility of recurrence after cancer treatment and improving the patient’s treatment plan.

Advantages of Sanger Sequencing Diagnostics

Sanger sequencing can detect mutant allele frequencies of about 10% to 20%.

Variant frequency (VAF) must be at least 10% to be reliable when using the Sanger sequencing method.

(Locked Nucleic Acid Technology for Highly Sensitive Detection of Somatic Mutations in Cancer, Takayuki Ishige, Sakae Itoga, Kazuyuki Matsushita (2018))

Application of ultra-precision restriction enzymes (GeneCker-Cas9, gcCas9)

With ultra-precision CRISPR/Cas (gcCas9) developed by GeneCker, CRISmono™ can be diagnosed at least 0.25% by overcoming the detection limit of existing Sanger Sequencing (<10%).

CRISmono™ Key features

Sanger Sequencing-based mutation detection kit

  • Gene mutation detection kit for high-performance liquid biopsy developed for MRD testing and diagnostic monitoring
 

Improved LoD (Limit of Detection) of genetic mutations with the use of ultra-precision genetic scissors

  • Application of ultra-precision CRISPR/Cas9 (gcCas9) system
  • Mutant gene amplification after the removal of a normal gene
 

Convenience of diagnosis

  • Test using blood
  • Suitable for periodic diagnostic monitoring with safe inspection
 

Gene customization

  • Customization for customer’s preference

CRISmono™ Working Process

Genomic DNA preparation & Non-target DNA removal step by gcCas9

CRISmono™, mutation detection kit for Sanger sequencing with ultra-precision CRISPR gene scissors (gcCas9) developed by GeneCker. gcCas9 is a tool for detecting cancer cell-derived mutation genes present in the blood, specifically cutting only normal genes and not affecting target mutation genes. Only target cancer genes can be detected through subsequent experiments.

PCR enrichment & DNA purification

Mutant genes that are cut and removed by gcCas9 must be amplified using PCR to obtain sufficient amounts. The mutant genes amplified through the PCR experimental process are purified using various DNA purification kits.

Sanger Sequencing & Analysis

Purified DNA can be sequenced through Sanger sequencing equipment. Due to the nature of the Sanger sequencing equipment, the equipment platform isn’t limited, and the results can be checked through the Sanger sequencing image viewer used in the laboratory.

CRISPincette™ NRAS

Sanger Sequencing-based mutation detection kit

NRAS mutation detection kit for high-performance Sanger Sequencing developed for NRAS gene’s Minimal Residual Disease (MRD) test and diagnostic monitoring.

Improved performance compared to existing Sanger Sequencing

For accurate detection of NRAS mutations, ultra-precision CRISPR/Cas9 (gcCas9) was applied, improving the detection performance of mutant genes by about 40 times compared to the existing Sanger Sequencing.

Product Specification

Product Specification
Input DNA Genomic DNA from blood ≥ 20 ng
Technology PCR Amplicon-Based
GeneCker - CRISPR/Cas9 (gcCas9)
Platform Sanger sequencer (ABI Genetic Analyzer)
Target Gene NRAS (G12R, G12D, G12A, G13D)
Target Size 221 bp
Limit of Detection, LoD ≥ 0.25% VAF (Mutation rate)
Target Gene mutation
NRAS G12R c.34G>C
G12D c.35G>A
G12A c.35G>C
G13D c.38G>A