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Journal articles
C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system
Philippe Jaïs
1, *
Etienne Decroly
2
Eric Jacquet
3
Marine Le boulch
1
Aurélien Jaïs
1
Olivier Jean-Jean
4
Heather Eaton
5
Prishila Ponien
3
Frédérique Verdier
6
Bruno Canard
2
Sergio Gonçalves
1
Stéphane Chiron
1
Maude Le Gall
7
Patrick Mayeux
6
Maya Shmulevitz
5
*
Corresponding author
Philippe H Jaïs 1
Correspondent author
Etienne Decroly 2
Author IdHAL : etienne-decroly ORCID : https://orcid.org/0000-0002-6046-024X
Eric Jacquet 3
Author
Marine Le boulch 1
Author
Frédérique Verdier 6
Author
Bruno Canard 2
Author IdHAL : bruno-canard ORCID : https://orcid.org/0000-0003-4924-1991
Sergio Gonçalves 1
Author
Stéphane Chiron 1
Author
1
Génopole Campus - Eukarÿs SAS [Evry, France] (France)
2
AFMB - Architecture et fonction des macromolécules biologiques (Faculté des Sciences de Luminy - ESIL 163 avenue de Luminy CP 925 13288 MARSEILLE CEDEX 09 - France)
- CNRS - Centre National de la Recherche Scientifique : UMR7257 (France)
- AMU - Aix Marseille Université : UMR7257 (Aix-Marseille Université Jardins du Pharo 58 Boulevard Charles Livon 13284 Marseille cedex 7 - France)
- INRA - Institut National de la Recherche Agronomique : USC1408 (France)
3
ICSN - Institut de Chimie des Substances Naturelles (Avenue de la terrasse 91198 Gif-sur-Yvette cedex - France)
- CNRS - Centre National de la Recherche Scientifique : UPR2301 (France)
4
B2A - Adaptation Biologique et Vieillissement = Biological Adaptation and Ageing (Campus de Jussieu - 4 place Jussieu 75005 Paris - France)
- INSERM - Institut National de la Santé et de la Recherche Médicale : U1164 (101, rue de Tolbiac, 75013 Paris - France)
- SU - Sorbonne Université (21 rue de l’École de médecine - 75006 Paris - France)
- CNRS - Centre National de la Recherche Scientifique : UMR8256 (France)
- IBPS - Institut de Biologie Paris Seine (Quai Saint-Bernard
75252 PARIS CEDEX 05 - France)
- INSERM - Institut National de la Santé et de la Recherche Médicale (101, rue de Tolbiac, 75013 Paris - France)
- SU - Sorbonne Université (21 rue de l’École de médecine - 75006 Paris - France)
- CNRS - Centre National de la Recherche Scientifique : FR 3631 (France)
5
Medical Microbiology and Immunology [Alberta, Canada] (6-142J Katz Group Centre for Pharmacy & Health Research - Edmonton, Alberta, T6G 2E1. - Canada)
- University of Alberta (116 St & 85 Ave, Edmonton, AB T6G 2R3 - Canada)
6
IC UM3 (UMR 8104 / U1016) - Institut Cochin (22 rue Méchain, 75014 Paris - France)
- INSERM - Institut National de la Santé et de la Recherche Médicale : U1016 (101, rue de Tolbiac, 75013 Paris - France)
- UPD5 - Université Paris Descartes - Paris 5 : UM 3 (12, rue de l'École de Médecine - 75270 Paris cedex 06 - France)
- CNRS - Centre National de la Recherche Scientifique : UMR 8104 (France)
7
CRI (UMR_S_1149 / ERL_8252 / U1149) - Centre de recherche sur l'Inflammation (Faculté de Médecine site Bichat - 16, Rue Henri Huchard - BP 416/4ème étage - 75870 Paris Cedex 18 - France)
- INSERM - Institut National de la Santé et de la Recherche Médicale : U1149 (101, rue de Tolbiac, 75013 Paris - France)
- CNRS - Centre National de la Recherche Scientifique : ERL_8252 (France)
- UPD7 - Université Paris Diderot - Paris 7 : UMR_S_1149 (5 rue Thomas-Mann - 75205 Paris cedex 13 - France)
Abstract : Most eukaryotic expression systems make use of host-cell nuclear transcriptional and post-transcriptional machineries. Here, we present the first generation of the chimeric cytoplasmic capping-prone phage polymerase (C3P3-G1) expression system developed by biological engineering, which generates capped and polyadenylated transcripts in host-cell cytoplasm by means of two components. First, an artificial single-unit chimeric enzyme made by fusing an mRNA capping enzyme and a DNA-dependent RNA polymerase. Second, specific DNA templates designed to operate with the C3P3-G1 enzyme , which encode for the transcripts and their artificial polyadenylation. This system, which can potentially be adapted to any in cellulo or in vivo eu-karyotic expression applications, was optimized for transient expression in mammalian cells. C3P3-G1 shows promising results for protein production in Chinese Hamster Ovary (CHO-K1) cells. This work also provides avenues for enhancing the performances for next generation C3P3 systems.
Document type :
Journal articles
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https://www.hal.inserm.fr/inserm-02016614
Contributor : Maude Le Gall <>
Submitted on : Wednesday, May 29, 2019 - 10:43:56 AM
Last modification on : Saturday, July 3, 2021 - 12:26:54 AM
Contributor : Maude Le Gall <>
Submitted on : Wednesday, May 29, 2019 - 10:43:56 AM
Last modification on : Saturday, July 3, 2021 - 12:26:54 AM
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gkz069.pdf
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Identifiers
- HAL Id : inserm-02016614, version 1
- PUBMEDCENTRAL : PMC6412113
- PUBMED : 30726994
- DOI : 10.1093/nar/gkz069
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INSERM | SU-SCIENCES | IBPS | B2A | CNRS | UNIV-PARIS7 | INRA | ICSN | UNIV-PARIS5 | INC-CNRS | UNIV-AMU | USPC | SORBONNE-UNIVERSITE | INRAE | UNIV-PARIS | UP-SANTE | SU-TI | GS-CHIMIE | GS-HEALTH-DRUG-SCIENCES
Citation
Philippe Jaïs, Etienne Decroly, Eric Jacquet, Marine Le boulch, Aurélien Jaïs, et al.. C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system. Nucleic Acids Research, Oxford University Press, 2019, 47 (5), pp.2681-2698. ⟨10.1093/nar/gkz069⟩. ⟨inserm-02016614⟩
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