Detection of large deletion in human BRCA1 gene in human breast carcinoma MCF-7 cells by using DNA-Silver Nanoclusters

Methods Appl Fluoresc. 2017 Dec 13;6(1):015001. doi: 10.1088/2050-6120/aa8988.

Abstract

Here we describe a label-free detection strategy for large deletion mutation in breast cancer (BC) related gene BRCA1 based on a DNA-silver nanocluster (NC) fluorescence upon recognition-induced hybridization. The specific hybridization of DNA templated silver NCs fluorescent probe to target DNAs can act as effective templates for enhancement of AgNCs fluorescence, which can be used to distinguish the deletion of BRCA1 due to different fluorescence intensities. Under the optimal conditions, the fluorescence intensity of the DNA-AgNCs at emission peaks around 440 nm (upon excitation at 350 nm) increased with the increasing deletion type within a dynamic range from 1.0 × 10-10 to 2.4 × 10-6 M with a detection limit (LOD) of 6.4 × 10-11 M. In this sensing system, the normal type shows no significant fluorescence; on the other hand, the deletion type emits higher fluorescence than normal type. Using this nanobiosensor, we successfully determined mutation using the non-amplified genomic DNAs that were isolated from the BC cell line.

MeSH terms

  • BRCA1 Protein / genetics*
  • Base Sequence*
  • Biosensing Techniques
  • Breast Neoplasms / genetics
  • DNA / chemistry*
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Humans
  • MCF-7 Cells
  • Nanostructures / chemistry*
  • Sequence Deletion*
  • Silver / chemistry*

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • Fluorescent Dyes
  • Silver
  • DNA