Experimental validation of a predicted microRNA within human FVIII gene.

Hemophilia A is an X-linked bleeding disorder that occurs due to the deficiency of Factor VIII (FVIII) protein clotting activity. The mutations in the F8 gene, which encodes FVIII coagulating protein have been widely reviewed. However, there is a wide range of criteria that in addition to F8 gene mutations, different molecular mechanisms may be associated with hemophilia A. Various functions of FVIII could be related to the hypothetical small non-coding RNAs, located within the F8 gene sequence. Therefore, miRNAs that can post-transcriptionally regulate gene expression might confer susceptibility to developing hemophilia A. Here, we have selected a bioinformatically predicted hairpin structure sequence in the first intron of the F8 gene that has the potential to produce a real miRNA (named put-miR1). We tried to experimentally detect the predicted miRNA via RT-PCR following its precursor overexpression in HEK 293 cell lines. Despite the accuracy of miRNA prediction, according to the reliable bioinformatics studies, we couldn't confirm the existence of considered mature miRNA in transfected cells. We hope that through changing experimental conditions, designing new primers, or altering cell lines and expression vectors, the exogenous and endogenous expression of the predicted miRNA will be confirmed.

MicroRNA-Targeted Signaling Pathways in the Autism Spectrum Disorder: Implications for Early Detection and Targeted Therapy.

BACKGROUND & OBJECTIVE: Autism Spectrum Disorders (ASD) is known as a neurodevelopmental disorder showing communication impairments and unusual patterns of behavior. Presently, it seems that ASD frequency is on the increase. Therefore, diagnostic tools that help detect the disease in the early stages can be very useful in better management of the disease. Recent studies demonstrate that miRNAs as novel biomarkers can be used to find out the process and etiology of ASD by regulating various genes of multiple pathways. However, ASD associated pathway targeted by miRNA is still in infancy. METHODS: In this in silico study taking into consideration the importance of miRNAs, we reviewed bioinformatics databases for finding possible pathways and potential miRNAs related to selected pathways. RESULTS: The results displayed some prominent pathways involved in ASD, as well as some experimental and predicted miRNAs that may regulate targets associated with these pathways such as neuroactive ligand-receptor interaction, serotonergic synapse, calcium signaling pathway, cAMP Signaling Pathway, PI3K-Akt signaling pathway. CONCLUSION: This study showed that the identified miRNAs may be involved in ASD-related pathways and may be considered as a new diagnostic tool and provide potential targets for the treatment of ASD.