Design of Nano bio-sensor based on graphene oxide for detection of mutation F508del in CFTR gene, leading to Cystic fibrosis

Introduction and objectives: Cystic fibrosis (CF) is one of the most common autosomal recessive diseases of Caucasian population which is caused by mutations in the Cystic Fibrosis Trans membrane conductance Regulator (CFTR) gene. The most common mutation for CF is referred to ΔF508, with a frequency of 86.4%. Cystic fibrosis is a multiple organ disease, which was affected on many organs such as lung, liver, pancreas, intestine bones etc. In the present work, a new nano-biosensor based on graphene oxide (GO) and 5-carboxy fluorescein (FAM) labeled DNA introduced for detecting deletion mutation in cystic fibrosis.Materials and methods: The graphite flake was purchase from Merck. According modified hummers method it was oxidized with H2SO4, HNO3 and KMnO4, under 55.0 °C for 12 h. Oligonucleotide sequences used in this study are as follows: 1) fDNA: 5-carboxy fluorescein fluorophore labeled at the 5′ terminal (the sequence of CFTR gene without any mutation located at codon 508). 2) cDNA the sequence of complementary target. 3) mDNA: the sequence of mismatch DNA possessing 21 bases deletion mutation at exon18 of CFTR gene. All nucleotides were purchased from BIONEER Co. (South Korea).Results: fDNA was quenched by GO, while adding cDNA caused double-stranded DNA formed and fluorescence (FL) emission was recovered again. By the optimum parameters in hand, the sensitivity of the designed probe for the detection of cDNA was investigated using various concentration of cDNA. It was deduced that, mDNA was unable to joint with fDNA and pick it up from GO surface so, FL emission was remain quenched.Conclusion: A new Nano bio-sensor was introduced for detection of Cystic fibrosis by studding on mutation located at codon 508, ΔF508.

کلیدواژه ها:

Cystic fibrosis ، graphene oxide ، mutation ، Nano-biosensor