Skip to main content
. 2022 Apr 19;13:2138. doi: 10.1038/s41467-022-29822-3

Fig. 5. The 22-bulge controls the cleavage activity of DICER on human pre-miRNAs.

Fig. 5

a, b Human pre-miRNAs contain a bulge in different positions on their 3p-strand (a) or 5p-strand (b). The pre-miRNA sequences were folded using RNAfold, and the number of pre-miRNAs containing a bulge in positions 18–26 from their 5p-end was quantified. c The number 59 of pre-miRNA orthologs containing a 22-bulge was determined. d The structures and sequences of 22-bulge pre-miRNAs and their nobulge variants. e The in vitro DICER cleavage assays of the 22-bulge and nobulge variants of human pre-miRNAs. f The cleavage efficiency of DICER on the 22-bulge and nobulge pre-miRNAs shown in panel (d), n = 3 independent experiments. The cleavage efficiency of DICER was calculated as the ratio of the cleaved product at DC21 or DC22 to the original substrate. A two-tailed t-test calculated the p-values. The error bars were presented with 95% confidence intervals. g The accuracy of DICER cleavage was calculated as the ratio of the cleaved product at DC21 to that at different positions. h The structures and sequences of the 22-bulge pre-miRNAs and their SNP variants. i The in vitro DICER cleavage assays of the 22-bulge and their SNP pre-miRNA variants. j The accuracy of DICER cleavage was calculated as the ratio of the cleaved product at DC21 to that at different positions. k Confirmation of the DICER cleavages on the pre-miRNAs by 5p-end of 3p-miRNAs. The sequences of miRNAs resulting from the ectopically expressing pre-mir-216a and pre-mir-376a-2 were determined by NGS. Source data are provided as a Source Data file.