RESEARCH ARTICLE


Design of FRET Probes for SNP RS1006737, Related to Mood Disorder



Germano Orrù1, 2, *, Mauro Giovanni Carta3, Alessia Bramanti4
1 Department of Surgical Sciences, Molecular Biology Service (MBS), University of Cagliari, Cagliari, Italy
2 National Research Council of Italy, ISPA, Sassari, Italy
3 Department of Medical Sciences and Public Health, University of Cagliari, Cagliari, Italy
4 Istituto di Scienze Applicate e Sistemi Intelligenti, ISASI, Messina, Italy


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© 2018 Orrù et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Department of Surgical Sciences, University of Cagliari, Germano Orrù Ph.D, via Ospedale 54, 09124 Cagliari, Italy; Tel: +39 070 609-2568; E-mail: orru@unica.it


Abstract

Background:

Several studies have shown that the Single Nucleotide Polymorphism (SNP) in the CACAN1C gene, rs1006737, is related to different mood disorder illnesses, such as bipolar disorder and schizophrenia. Current day molecular procedures for allele detection of this gene can be very expensive and time consuming. Hence, a sensitive and specific molecular procedure for detecting these mutations in a large number of subjects is desirable, especially for research groups who have no complex laboratory equipment.

Objective:

The possibility of using a Fluorescence Resonance Energy Transfer (FRET) probe was evaluated by means of bioinformatic tools, designed for forecasting the molecular behavior of DNA probes used in the research field or for laboratory analysis methods.

Method:

In this study we used the DINAMelt Web Server to predict the Tms of FRET oligo in the presence of the A and/or G allele in rs1006737. The PCR primers were designed by using oligo 4 and oligo 6 primer analysis software,

Results:

The molecular probe described in this study detected a Tm difference of 5-6°C between alleles A and G in rs1006737, which also showed good discrimination for a heterozygous profile for this genomic region.

Conclusion:

Although in silico studies represent a relatively new avenue of inquiry, they have now started to be used to predict how a molecular probe interacts with its biological target, reducing the time and costs of molecular test tuning. The results of this study seem promising for further laboratory tests on allele detection in rs1006737 region.

Keywords: Mood disorders, Genome-wide association studies (GWAS), CACNA1C gene, RS1006737, FRET probes, Real time PCR.