Abstract
Peptide nucleic acids (PNAs) have gained much interest as molecular recognition tools in biology, medicine and chemistry. This is due to high hybridization efficiency to complimentary oligonucleotides and stability of the duplexes with RNA or DNA. We have synthesized 15/16-mer PNA probes to detect the HER2 mRNA. The performance of these probes to detect the HER2 target was evaluated by fluorescence imaging and fluorescence bead assays. The PNA probes have sufficiently discriminated between the wild type HER2 target and the mutant target with single base mismatches. Furthermore, the probes exhibited excellent linear concentration dependence between 0.4 to 400 fmol for the target gene. The results demonstrate potential application of PNAs as diagnostic probes with high specificity for quantitative measurements of amplifications or over-expressions of oncogenes.
Highlights
There is a growing demand for rapid methods of detection for genetic aberrations that are implicated in various diseases
We developed peptide nucleic acid (PNA) probes for the detection of an oncogene known as HER2, which encodes a 185 kDa tyrosine kinase in the family of human epidermal growth factor receptors
We synthesized several PNA probes targeting the HER2 RNA and in most cases the automated synthesis produced essentially full length PNA probes with good yields; after HPLC purification the products were characterized by mass spectrometry (Table 1)
Summary
There is a growing demand for rapid methods of detection for genetic aberrations that are implicated in various diseases. It has been shown that 20–30% of breast cancer patients have amplification and over-expression of HER2 oncogene which have been correlated with aggressive, drug resistant and poor prognosis in breast cancer [2] These characteristics clearly establish the potential use of HER2 as a biomarker of the disease as well as for the development of targeted and personalized treatment [3,4,5]. There are two FDA approved HER2 tests in the clinics namely; immuno-histochemistry (IHC) and fluorescence in-situ hybridization (FISH) which are mainly applied to strategize therapeutic regimen [6,7] Both tests are semi-quantitative and require sophisticated laboratory techniques and instrumentation, it is desirable to develop simpler, sensitive, rapid, and scaleable genetic detection methods to check the HER2 status of breast cancer patients [8,9]
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