PT  - JOURNAL ARTICLE
AU  - Mei-Mei Li
AU  - Balasubrahmanyam Addepalli
AU  - Mei-Juan Tu
AU  - Qiu-Xia Chen
AU  - Wei-Peng Wang
AU  - Patrick A. Limbach
AU  - Janine M. LaSalle
AU  - Su Zeng
AU  - Min Huang
AU  - Ai-Ming Yu
TI  - Chimeric MicroRNA-1291 Biosynthesized Efficiently in <em>Escherichia coli</em> Is Effective to Reduce Target Gene Expression in Human Carcinoma Cells and Improve Chemosensitivity
AID  - 10.1124/dmd.115.064493
DP  - 2015 Jul 01
TA  - Drug Metabolism and Disposition
PG  - 1129--1136
VI  - 43
IP  - 7
4099  - http://dmd.aspetjournals.org/content/43/7/1129.short
4100  - http://dmd.aspetjournals.org/content/43/7/1129.full
SO  - Drug Metab Dispos2015 Jul 01; 43
AB  - In contrast to the growing interests in studying noncoding RNAs (ncRNAs) such as microRNA (miRNA or miR) pharmacoepigenetics, there is a lack of efficient means to cost effectively produce large quantities of natural miRNA agents. Our recent efforts led to a successful production of chimeric pre-miR-27b in bacteria using a transfer RNA (tRNA)–based recombinant RNA technology, but at very low expression levels. Herein, we present a high-yield expression of chimeric pre-miR-1291 in common Escherichia coli strains using the same tRNA scaffold. The tRNA fusion pre-miR-1291 (tRNA/mir-1291) was then purified to high homogeneity using affinity chromatography, whose primary sequence and post-transcriptional modifications were directly characterized by mass spectrometric analyses. Chimeric tRNA/mir-1291 was readily processed to mature miR-1291 in human carcinoma MCF-7 and PANC-1 cells. Consequently, recombinant tRNA/mir-1291 reduced the protein levels of miR-1291 target genes, including ABCC1, FOXA2, and MeCP2, as compared with cells transfected with the same doses of control methionyl-tRNA scaffold with a sephadex aptamer (tRNA/MSA). In addition, tRNA-carried pre-miR-1291 suppressed the growth of MCF-7 and PANC-1 cells in a dose-dependent manner, and significantly enhanced the sensitivity of ABCC1-overexpressing PANC-1 cells to doxorubicin. These results indicate that recombinant miR-1291 agent is effective in the modulation of target gene expression and chemosensitivity, which may provide insights into high-yield bioengineering of new ncRNA agents for pharmacoepigenetics research.