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. 2018 Jun 15;7:e34268. doi: 10.7554/eLife.34268

Figure 4. RNA-RNA interaction between intron and mRNA.

Figure 4.

(A) Schematic of construct harboring RNA aptamer. Pre-mRNA bears the Ll.LtrB intron (red line), which is flanked by small exons (brown and orange boxes). The RNA streptavidin aptamer (SA) is shown as a green stem-loop. Splicing of the aptamer-containing (Int+SA+) or aptamer-less (Int+SA-) pre-mRNA produces small ligated exons (S-smRNA) that have the same sequence as the control small mRNA (C-smRNA) generated from the intron-less construct (Int-). Primers IDT5078 and IDT1073 were used for analysis of smRNAs and intron RNAs (pre-mRNA and spliced intron), respectively, in panel below and are indicated as arrows. (B) mRNA binds to the intron (representative result of three biological replicates). RNAs with the SA aptamer were purified using streptavidin resin and were detected by reverse transcription using 5′- 32P-labeled primers specific for smRNAs and intron RNAs (pre-mRNA and Intron). The cDNA products were resolved on an 8% urea-polyacrylamide gel. Cellular 6S non-coding RNA was used as a loading control. F, flow-through; W4, the fourth wash; W8, the eighth wash, B, resin-bound.