New Assays for Expression Profiling of MicroRNAs

DESCRIPTION provided by applicant MicroRNAs miRNAs are involved in many diverse biological processes and a single miRNA can modulate the expression of hundreds of gene targets Changes in miRNA expression could have profound biological impacts leading to a variety of human diseases To date most miRNA expression profiling studies have relied on microarrays or high throughput RNA sequencing RNA seq For these high throughput experiments it is important to validate the profiling results with independent methods Real time PCR is considered as the gold standard for gene expression quantification and thus is ideal for validation of microarray or RNA seq results Furthermore despite their wide use for miRNA profiling there are some major limitations associated with microarrays and RNA seq such as the requirement of high quality RNA samples To address these issues we have developed a new real time RT PCR method With this new method we have profiled the expression of cancer related miRNAs in hundreds of archived tumor tissues and identified novel miRNA signatures as predictive biomarkers for multiple types of cancer Building on the success of our new method for cancer related miRNA profiling we propose to significantly expand the scope of the method by designing and testing new assays to encompass all known miRNAs in the human genome Specific Aim These miRNA assays will serve two main purposes As a reliable independent method for validation of high throughput results from microarrays or RNA seq and expression profiling of low quality degraded RNA extracted from clinical tissues In both cases availability of these miRNA assays will enable researchers to correlate abnormal miRNA expression changes to human diseases by focusing on a selected set of miRNAs To facilitate the selection of miRNA assays by end users we also propose to establish an online database to present experimentally validated assays for all known human miRNAs Specific Aim PUBLIC HEALTH RELEVANCE MicroRNAs miRNAs are involved in many diverse biological processes and changes in miRNA expression lead to various human diseases One major challenge in this field is the lack of assays for miRNA expression profiling analysis To address this issue we propose to develop new assay products to profile miRNA expression changes related to human diseases