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

Small RNA sequencing of all the let-7a single-nucleotide mutants to confirm the expression of the mutant miRNAs.We found that the abundance of the mutated let-7a miRNA in each mutant was similar to that of the native let-7a in WT, with a variance (<10%) that we judge not sufficient to cause lf phenotypes by reduced expression (Fig. 3B). Note that the strains with mutations at g11-g16 also contained a WT let-7 allele on a genetic balancer umnIs25(mnDp1), hence both WT and mutant let-7a were expressed. Overall design: small RNA seq of let-7a 3' non-seed single nucleotide mutants.