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High Throughput miRNA Screening Identifies miR-574-3p Hyperproductive Effect in CHO Cells
Živa Švab1, Luca Braga2, Corrado Guarnaccia1
1Biotechnology Development Unit, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
Biomolecules
|August 27, 2021
Summary
We identified specific microRNAs (miRNAs) that significantly boost the production of biotherapeutics in CHO cells. Overexpressing miR-574-3p increased Erythropoietin and Etanercept yields by up to two-fold.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Chinese Hamster Ovary (CHO) cells are crucial for manufacturing complex biotherapeutics.
- Increasing cell productivity is essential to meet the rising demand for these drugs.
- MicroRNA (miRNA) manipulation offers a powerful strategy for enhancing cellular pathways without significant translational burden.
Purpose of the Study:
- To screen human miRNAs for their ability to enhance biotherapeutic production in CHO cells.
- To identify specific miRNAs that increase Erythropoietin and Etanercept titers.
- To investigate the mechanism of miRNA-mediated productivity enhancement in CHO cells.
Main Methods:
- High-throughput screening of 2042 human miRNAs in CHO cells.
- Overexpression of candidate miRNAs and measurement of Erythropoietin and Etanercept production.
- Target validation for identified miRNAs, including p300.
- Comparison of miRNA expression using human versus endogenous CHO pri-miRNA sequences.
Main Results:
- Several human miRNAs were identified that increase Erythropoietin and Etanercept production in CHO cells.
- miR-574-3p overexpression resulted in a 1.3 to 2-fold increase in biotherapeutic titers.
- p300 was validated as a key target of miR-574-3p in CHO cells.
- CHO cells engineered with endogenous pri-miRNA sequences showed superior miRNA expression compared to those with human sequences.
Conclusions:
- miRNA engineering is a viable strategy for enhancing biotherapeutic production in CHO cells.
- miR-574-3p is a potent miRNA for improving Erythropoietin and Etanercept yields.
- The flanking genomic sequences and secondary structure of pri-miRNAs significantly influence their processing and efficacy.
- This study provides a novel, efficient method for miRNA engineering in CHO cells.
