Engineering mammalian cell factories with SINEUP noncoding RNAs to improve translation of secreted proteins

Gene. 2015 Sep 15;569(2):287-93. doi: 10.1016/j.gene.2015.05.070. Epub 2015 Jun 2.

Abstract

Whenever the function of a recombinant protein depends on post-translational processing, mammalian cells become an indispensable tool for their production. This is particularly true for biologics and therapeutic monoclonal antibodies (MAbs). Despite some drawbacks, Chinese Hamster Ovary (CHO) cells are the workhorse for MAbs production in academia and industry. Several methodologies have been adopted to improve expression and stability, including methods based on selective pressure or cell engineering. We have previously identified SINEUPs as a new functional class of natural and synthetic long non-coding RNAs that through the activity of an inverted SINEB2 element are able to promote translation of partially overlapping sense coding mRNAs. Here we show that by taking advantage of their modular structure, synthetic SINEUPs can be designed to increase production of secreted proteins. Furthermore, by experimentally validating antisense to elastin (AS-eln) RNA as a natural SINEUP, we show that SINEUP-mediated control may target extracellular proteins. These results lead us to propose synthetic SINEUPs as new versatile tools to optimize production of secreted proteins in manufacturing pipelines and natural SINEUPs as new regulatory RNAs in the secretory pathways.

Keywords: CHO cell; Cell factory; Elastin; Long noncoding RNA; Luciferase assay; SINEUP; Serum-free; Signal peptide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CHO Cells
  • Cell Adhesion Molecules / genetics
  • Cell Engineering*
  • Cricetulus
  • Elastin / genetics
  • Humans
  • Molecular Sequence Data
  • Protein Biosynthesis*
  • RNA, Antisense / chemistry
  • RNA, Antisense / genetics
  • RNA, Antisense / metabolism*
  • RNA, Long Noncoding / chemistry
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • Recombinant Proteins / metabolism*
  • Secretory Pathway

Substances

  • Cell Adhesion Molecules
  • POSTN protein, human
  • RNA, Antisense
  • RNA, Long Noncoding
  • Recombinant Proteins
  • Elastin