Novel RNA molecular bioengineering technology efficiently produces functional miRNA agents

Comparison of computationally predicted 3D structures of BioRNAs. (A) Predicted 3D structures of human leucyl (UAA) and glycyl (GCC) tRNAs, hsa-pre-miR-34a, and the hybrid carrier RNAs (BioRNA^Gly/miR-34a-5p and BioRNA^Leu/miR-34a-5p). (B) Predicted structures of the hsa-miR-7 duplex, hsa-pre-miR-34a carrier with payload miR-7 (hsa-pre-miR-34a/miR-7-5p), BioRNA^Gly/miR-7-5p, and BioRNA^Leu/miR-7-5p. (C) 3D structures of the hsa-miR-124 duplex, hsa-pre-miR-34a carrier substituted by miR-124 duplex (hsa-pre-miR-34a/miR-124-3p), BioRNA^Gly/miR-124-3p, and BioRNA^Leu/miR-124-3p. Colored subcomponents of BioRNAs consist of ribbon structures: htRNA (black), hsa-pre-miR-34a (blue), miRNA guide (antisense; red) and passenger (sense; light blue or light gray) sequence, five-prime (5′; magenta), and three-prime (3′; white). Nucleotides consist of color-filled nucleobases: adenine (red), thymine (blue), guanine (green), and cytosine (yellow). Modeling results demonstrate a retained overall structure of BioRNAs when substituted with target miRNAs. All RNA 3D structures were generated by using RNAComposer after inputting the secondary structures predicted by RNAFold, based on the primary sequences shown in Supplemental Tables S1 and S4. The resulting structures were adapted using UCSF ChimeraX next-generation molecular visualization program.