title: Engineering mammalian cell factories with SINEUP noncoding RNAs to improve translation of secreted proteins.
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.
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Patrucco
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Chiesa
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy.
authors.forename: Maria Felicia
authors.initials: MF
authors.lastname: Soluri
authors.affiliation: Area of Neuroscience, SISSA, Trieste, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Fasolo
authors.affiliation: RIKEN Center for Life Science Technologies, Division of Genomic Technologies, Yokohama, Japan.
authors.forename: Hazuki
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: RIKEN Center for Life Science Technologies, Division of Genomic Technologies, Yokohama, Japan; TransSINE Technologies, Yokohama, Japan.
authors.forename: Piero
authors.initials: P
authors.lastname: Carninci
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy; Area of Neuroscience, SISSA, Trieste, Italy; TransSINE Technologies, Yokohama, Japan.
authors.forename: Silvia
authors.initials: S
authors.lastname: Zucchelli
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy; TransSINE Technologies, Yokohama, Japan.
authors.forename: Claudio
authors.initials: C
authors.lastname: Santoro
authors.affiliation: Area of Neuroscience, SISSA, Trieste, Italy; TransSINE Technologies, Yokohama, Japan.
authors.forename: Stefano
authors.initials: S
authors.lastname: Gustincich
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy; University of Trieste, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Sblattero
authors.affiliation: Department of Health Science and Interdisciplinary Research Center on Autoimmune Diseases, Università del Piemonte Orientale, Novara, Italy. Electronic address: diego.cotella@med.uniupo.it.
authors.forename: Diego
authors.initials: D
authors.lastname: Cotella
mesh_terms: D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000235:genetics; D060846:Cell Engineering; D003412:Cricetulus; D004549:Elastin / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D062085:RNA, Long Noncoding / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D055571:Secretory Pathway
chemical_list: D015815:Cell Adhesion Molecules; C512835:POSTN protein, human; D016372:RNA, Antisense; D062085:RNA, Long Noncoding; D011994:Recombinant Proteins; D004549:Elastin
keywords: CHO cell; Cell factory; Elastin; Long noncoding RNA; Luciferase assay; SINEUP; Serum-free; Signal peptide