Zucchelli, E;
Pema, M;
Stornaiuolo, A;
Piovan, C;
Scavullo, C;
Giuliani, E;
Bossi, S;
... Bovolenta, C; + view all
(2017)
Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein.
Molecular Therapy - Methods and Clinical Development
, 4
pp. 102-114.
10.1016/j.omtm.2017.01.002.
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Abstract
Lentiviral vectors (LVs) are a highly valuable tool for gene transfer currently exploited in basic, applied, and clinical studies. Their optimization is therefore very important for the field of vectorology and gene therapy. A key molecule for LV function is the envelope because it guides cell entry. The most commonly used in transiently produced LVs is the vesicular stomatitis virus glycoprotein (VSV-G) envelope, whose continuous expression is, however, toxic for stable LV producer cells. In contrast, the feline endogenous retroviral RD114-TR envelope is suitable for stable LV manufacturing, being well tolerated by producer cells under constitutive expression. We have previously reported successful, transient and stable production of LVs pseudotyped with RD114-TR for good transduction of T lymphocytes and CD34+ cells. To further improve RD114-TR-pseudotyped LV cell entry by increasing envelope expression, we codon-optimized the RD114-TR open reading frame (ORF). Here we show that, despite the RD114-TRco precursor being produced at a higher level than the wild-type counterpart, it is unexpectedly not duly glycosylated, exported to the cytosol, and processed. Correct cleavage of the precursor in the functional surface and transmembrane subunits is prevented in vivo, and, consequently, the unprocessed precursor is incorporated into LVs, making them inactive.
| Type: | Article |
|---|---|
| Title: | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.omtm.2017.01.002 |
| Publisher version: | https://doi.org/10.1016/j.omtm.2017.01.002 |
| Language: | English |
| Additional information: | This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Medicine, Research & Experimental, Research & Experimental Medicine, Simian Immunodeficiency Virus, Packaging Cell-Line, Lentiviral Vectors, Messenger-Rna, Synonymous Mutations, Usage Bias, Secondary Structure, Protein Expression, Escherichia-Coli, Gene-Expression |
| UCL classification: | UCL |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/1550684 |
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