Cargando…
Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein
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...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5363313/ https://www.ncbi.nlm.nih.gov/pubmed/28344996 http://dx.doi.org/10.1016/j.omtm.2017.01.002 |
_version_ | 1782517138957795328 |
---|---|
author | Zucchelli, Eleonora Pema, Monika Stornaiuolo, Anna Piovan, Claudia Scavullo, Cinzia Giuliani, Erica Bossi, Sergio Corna, Stefano Asperti, Claudia Bordignon, Claudio Rizzardi, Gian-Paolo Bovolenta, Chiara |
author_facet | Zucchelli, Eleonora Pema, Monika Stornaiuolo, Anna Piovan, Claudia Scavullo, Cinzia Giuliani, Erica Bossi, Sergio Corna, Stefano Asperti, Claudia Bordignon, Claudio Rizzardi, Gian-Paolo Bovolenta, Chiara |
author_sort | Zucchelli, Eleonora |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5363313 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-53633132017-03-24 Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein Zucchelli, Eleonora Pema, Monika Stornaiuolo, Anna Piovan, Claudia Scavullo, Cinzia Giuliani, Erica Bossi, Sergio Corna, Stefano Asperti, Claudia Bordignon, Claudio Rizzardi, Gian-Paolo Bovolenta, Chiara Mol Ther Methods Clin Dev Original Article 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. American Society of Gene & Cell Therapy 2017-01-11 /pmc/articles/PMC5363313/ /pubmed/28344996 http://dx.doi.org/10.1016/j.omtm.2017.01.002 Text en © 2017 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Zucchelli, Eleonora Pema, Monika Stornaiuolo, Anna Piovan, Claudia Scavullo, Cinzia Giuliani, Erica Bossi, Sergio Corna, Stefano Asperti, Claudia Bordignon, Claudio Rizzardi, Gian-Paolo Bovolenta, Chiara Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title_full | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title_fullStr | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title_full_unstemmed | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title_short | Codon Optimization Leads to Functional Impairment of RD114-TR Envelope Glycoprotein |
title_sort | codon optimization leads to functional impairment of rd114-tr envelope glycoprotein |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5363313/ https://www.ncbi.nlm.nih.gov/pubmed/28344996 http://dx.doi.org/10.1016/j.omtm.2017.01.002 |
work_keys_str_mv | AT zucchellieleonora codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT pemamonika codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT stornaiuoloanna codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT piovanclaudia codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT scavullocinzia codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT giulianierica codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT bossisergio codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT cornastefano codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT asperticlaudia codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT bordignonclaudio codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT rizzardigianpaolo codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein AT bovolentachiara codonoptimizationleadstofunctionalimpairmentofrd114trenvelopeglycoprotein |