E-MTAB-1612 - CLIP-Seq and RNA-Seq of liver samples from untreated mice and those which have undergone partial hepatetomy to study how miRNAs are recruited to mRNA targets in regenerating liver
Released on 25 April 2013, last updated on 3 May 2014
Validation of physiologic miRNA targets has been met with significant challenges. We employed HITS-CLIP to identify which miRNAs participate in liver regeneration, and to identify their target mRNAs. miRNA recruitment to the RISC is highly dynamic, changing more than five-fold for several miRNAs. miRNA recruitment to the RISC did not correlate with changes in overall miRNA expression for these dynamically recruited miRNAs, emphasizing the necessity to determine miRNA recruitment to the RISC in order to fully assess the impact of miRNA regulation. We incorporated RNAseq quantification of total mRNA to identify expression-weighted Ago footprints, and developed a microRNA regulatory element (MRE) prediction algorithm that represents a greater than 20-fold refinement over computational methods. These high confidence MREs were used to generate candidate competing endogenous RNA (ceRNA) networks. Conclusion: HITS-CLIP analysis provide novel insights into global miRNA:mRNA relationships in the regenerating liver .
CLIP-Seq, RNA-seq of coding RNA, co-expression, in vivo, stimulus or stress, time series
Jonathan Schug <firstname.lastname@example.org>, A Vourekas, Gabriel Walton, Joshua R Friedman, K Essuman, Karen Markley, Klaus H Kaestner <email@example.com>, Linda E Greenbaum <firstname.lastname@example.org>, Lindsey B McKenna, M Nakagawa, Nicholas J Hand, S Mukherjee, Vasumathi Kameswaran, Y Gao, Z Shi
Dynamic Recruitment of microRNAs to their mRNA Targets in the Regenerating Liver. Schug, J., McKenna, L.B., Walton, G., Hand, N.J., Mukherjee, S., Essuman, K., Shi, Z., Gao, Y., Markley, K., Nakagawa, M., Kameswaren, V., Vourekas, A., Friedman, J.R., Kaestner, K.H., and L.E. Greenbaum.