Epigenetics

Effects of Cadmium on the epigenetics of cytoskeleton genes in CHO cellsEpigenetics and Heavy Metal Homeostasis in Arabidopsis Thaliana

Idalisse Colón Rodríguez, B.S.
Juan A. Negrón Berríos, Ph.D.
Interamerican University of Puerto Rico
Barranquitas Campus

Cadmium is an environmental contaminant that causes cell damage. The term epigenetic refers to changes in gene expression that do not involve changes in the DNA base sequence, but rather DNA methylation and histone alterations. This study focuses on investigating the epigenetics of cytoskeletal genes in CHO cell cultures when exposed to cadmium. In particular, we are interested in investigating the epigenetics of actin and myosin when exposed to highly toxic conditions. The epigenetics associated with DNA methylation in cells exposed and not exposed to cadmium was compared. To determine epigenetic changes in these cells, measurements of methylation levels were used as provided by the “Melting Curve Analysis”. Genomic DNA was isolated and converted by the bisulfate reaction. Bioinformatics analysis was used to develop primers specific for the promoter of the actin and myosin region of CHO cells. Results are in progress to determine whether or not these genes are epigenetically affected in the presence of cadmium.

Idalisse Colón Rodríguez, B.S.
Juan A. Negrón Berríos, Ph.D.
Interamerican University of Puerto Rico
Barranquitas Campus

mRNAs are non-coding RNA molecules 21-23 nucleotides long, have been shown to have regulatory effects on gene expression. The central purpose of this research is to study the expression of miRNA in the Arabidopsis thaliana plant when exposed to cadmium. Cadmium is a highly toxic metal for most plants, animals and humans. In this research we want to analyze the possible relationship between non-coding RNAs and the plant response to stress induced by heavy metals. The in vitro cultures of Arabidopsis thaliana exposed to cadmium, the total RNA was extracted, and separated in a denaturing polyacrylamide gel. Bands corresponding to the expected miRNA size were cut out and extracted. The concatamers were converted to cDNA and cloned into an appropriate vector. By sequencing the generated clones, several miRNAs were identified, such as osa-miR-1425-5p. Future studies are necessary to establish a clear relationship between these miRNAs and the plant response when exposed to cadmium-induced stress.