Critical contribution of 3' non-seed base pairing to the in vivo function of the evolutionarily conserved let-7a microRNA [RNA-seq]

The C.elegans transcriptomes of single-nt let-7a 3' non-seed mutants (U13A / U18A). In order to identified gene perturbance and determine the post-transcriptional regulation mechanism caused by disrupting 3' non-seed pairing of let-7a, we examined the global gene expression by RNAseq in addition to ribosome profiling of L4 whole animals. To include the deadenylated but temperally stable mRNA in the enrichment, we used a RNase H based ribosomal RNA depletion with antisense oligos specifically designed for C.elegans to enrich mRNA (PMID: 33005886). lin-41(ma480) was used in the background to filter out downstream regulation related to the lin-41 pathway. Overall design: RNA-seq of L4 larvae of DG3913[lin-41(tn1541)], VT3878[lin-41(tn1541ma480);let-7(ma432)], VT1367(maIs105), VT3797[ma105,let-7(ma435)] at 20 °C.