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Effects of Cadmium on Epigenetics of Cytoskeletal Genes in CHO Cells
Author(s) -
Colón Rodríguez Idalisse,
Negrón Berrios Juan
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.884.47
Subject(s) - dna methylation , epigenetics , myosin , cytosine , microbiology and biotechnology , bisulfite sequencing , chinese hamster ovary cell , methylation , actin , dna , biology , gene , cpg site , cytoskeleton , chemistry , biochemistry , gene expression , genetics , cell , cell culture
Epigenetic is a mechanism that involves changes in the expression of genes, which are not involved in changes or alterations in the sequence of the DNA strands. DNA methylation is one of the most studied chemical changes associated to epigenetics. Highly methylated DNA is usually associated to inhibition of transcriptions. Actin cytoskeleton provides support and internal organization to the cell. It is also involved in intracellular transport. The goal of this study is to investigate epigenetics of β actin non‐muscle myosin type II in cytoskeleton of Chinese Hamster Ovaries cells (CHO cells) exposed to cadmium. In particular, we are interested in the methylation profile in β actin and non‐muscle myosin type II during the toxic conditions induced by the heavy metal. Genomic DNA was isolated and treated with the bisulfite reaction, in which the un‐methylated cytosine were transformed in thymine, later using PCR reaction these thymine residues were transformed in uracil, whereas, the methylated cytosine residues remain unchanged. High Resolution Melting (HRM) analysis through real time PCR assay was used to obtain the percentage of DNA methylation. This allows us to discriminate between methylated and non‐methylated cytosine. Bioinformatics was used to develop specific primers for the promoter region of actin and myosin from CHO cells. Primers were calibrated using regular and real time PCR, to make sure of their specificity. Preliminary results indicate differences in methylation profile at the promoter region of actin and myosin genes in CHO cells during to exposition to, cadmium.