Bioinformatics study and cloning of a stem loop structure required for it's over expression

ERBB2 is a member of the human epidermal growth factor receptor family and is located at large arm of chromosome 17. This receptor can activate a variety of signaling pathways which inhibit apoptosis and induce cell proliferation. MiRNAs are small, single strand, non coding RNAs that regulate gene expression at post transcriptional level and control important cellular processes such as proliferation, differentiation, apoptosis and tumor genesis. Genomic location of miRNAs demonstrated that they are located at fragile and/ or cancer related chromosomal regions. The aim of this study is production of necessary structure to over express the predicted area of a novel miRNA in ERBB2 gene following its bioinformatics prediction.

Different bioinformatics software was used to investigate stem loop structures with a potential to produce novel miRNAs, Dicer and Drosha enzymes recognition sites and conservation of interested area. Then the surrounding area of candidate stem loop structure was cloned in pEGFPC1 expression vector successfully.

Different software study shows a high probability of processing a mature miRNA. The required construct for more studies was cloned in pEGFPC1 expression vector successfully.

Bioinformatics analysis predictING stem loop structure has a potential to produce a novel miRNA.