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. 2019 Nov 7;10(1):159–170. doi: 10.1016/j.apsb.2019.11.001

Figure 1.

Figure 1

Production of fully-humanized biologic miR-328-3p molecule. (A) Schematic illustration of the fully-humanized bioengineered RNA agent (hBERA) bearing the target miR-328-3p (hBERA/miR-328, 192 nucleotides in length). The carrier consists of a human serine tRNA (htRNASer) and a modified pre-miR-34a23, in which miR-34a duplexes are replaced by target miR-328 sequences. (B) High-level heterogeneous expression of hBERA/miR-328 through bacterial fermentation, as demonstrated by urea-PAGE analysis. Total RNA from wild type (WT) Escherichia coli was used for comparison. (C) FPLC trace during the purification of hBERA/miR-328. The insert shows urea-PAGE analysis of target RNA fractions collected during FPLC separation. (D) The purity of isolated hBERA/miR-328-3p (98.5%) was determined by HPLC analysis. Au/Uv, absorbance unit of ultraviolet-visible spectroscopy; nt, nucleotide.