A comprehensive in silico approach for finding novel microRNAs in human genome. Novel transcipt in ZC3HAV1 gene

By using computational analysis of the last exon of the ZC3HAV1 gene we found a hairpin-shaped segment which potentially could be a microRNA. For this reason, we predicted the putative mature sequence of the hairpin and then a primer was designed followed by RT-PCR amplification together with a universal Reverse primer. As predicted, a PCR product with about 80 nucleotides long fragment was observed in a colorectal cancer-originated cell line (SW480). Cloning this PCR product in TA vector with subsequent Sanger sequencing method confirmed its endogenous expression in this cell line. The resultant sequence is unique in human genome and it completely matches to the last exon of the studied gene. This is the first report of the existence of a novel microRNA located in this genome region.