Genome-wide Direct Transcription Measurement using Array-based Assay of Labeled Nascent RNA
Description (Set) JHU scientists have developed, optimized and validated an innovative approach for study of nascent RNA transcription. Post-transcriptional regulation of gene expression constitutes critical and wide-spread mechanisms. However, only ongoing transcription accurately reflects momentary regulatory events of transcriptional regulatory mechanisms. Ongoing transcription can be measured and distinguished from mRNA sequence turnover using Nuclear run-on (NRO) assays. Traditional NRO assays have several limitations due to metabolic labeling of new RNA with a radioactive nucleotide. JHU scientists have developed an array-based assay that is an improvement over existing methods because it non-radioactively labels newly transcribed mRNA and can be used for genome wide transcription studies. This method combines classical NRO techniques with standard conventional microarray methods. Benefits: ? Drastically reduced cell numbers required for starting material. ? Applicable both in vitro and in vivo. ? Efficient and specific selection of labeled nascent RNA. ? Detection of all processed mRNA including non-coding RNA. ? Analysis of labeled newly transcribed RNA at a full genome scale. ? Direct link microRNA expression detection with changes in the expression of genes whose regulation they control. Proposed Use (Set) This technology could be commercialized as labeling reagents for newly transcribed RNA and specific arrays for nascent RNA detection. It could also be commercialized as an assay to study transcription and post-transcriptional regulation of multiple RNA species including microRNA, antisense mRNA and other non-coding RNA.
Fan, Jinshui ,Barnes, Kathleen,Cheadle, Chris,Chen, Yu-chi,Watkins, Tonya
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