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Synthetic transactivator–promoter systems for exogenous gene expression in Chlamydomonas reinhardtii.

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Title: Synthetic transactivator–promoter systems for exogenous gene expression in Chlamydomonas reinhardtii.
Authors: Kawabe, Yoshinori1 (AUTHOR) kawabe@chem-eng.kyushu-u.ac.jp, Akiyama, Tatsuki1 (AUTHOR), Miyazoe, Kokoro1 (AUTHOR), Kamihira, Masamichi1 (AUTHOR)
Source: Journal of Bioscience & Bioengineering. Jun2026, Vol. 141 Issue 6, p430-437. 8p.
Subjects: Transgene expression, Genetic transcription regulation, Transcription factors, Protein expression, Chlamydomonas reinhardtii
Abstract: Chlamydomonas reinhardtii , which is a unicellular photosynthetic eukaryote, has long served as a model microalga for fundamental biological research and biotechnological applications. Recently, it has attracted attention as a promising biological resource for the sustainable production of bio-oils and high-value biomolecules. To enhance the biotechnological utility of this species, various genetic engineering tools have been developed in recent years. In this study, the tetracycline repressor (TetR)-based transactivation system, which is widely used in mammalian and other eukaryotic cells, was repurposed to establish a synthetic transcriptional activation system for exogenous gene expression in Chlamydomonas. We first constructed a transient expression-based evaluation platform to screen for transcriptional activation domains (TADs) that are functional in Chlamydomonas. Among the analyzed TADs, VP192 (tandem repeat of 12 copies of the core VP16 domain) had the highest transcriptional activity when fused to either TetR or Gal4 DNA-binding domains. Furthermore, fusion proteins comprising VP192 and either TetR or reverse TetR enabled doxycycline-dependent regulation of transgene expression in a dose-dependent manner. Notably, transcriptional inducibility was maintained even when the tetracycline-responsive element (TRE) was fused to the HSP70A promoter. Combining TetR-VP192 and this synthetic chimeric promoter (TRE-P HSP) yielded transgene expression levels that exceeded those resulting from the strong HSP70A / RbcS2 hybrid promoter by more than 14-fold. These findings suggest that artificial transcription factors and engineered promoters provide a versatile molecular toolkit for regulating exogenous gene expression in Chlamydomonas. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Bioscience & Bioengineering is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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DbLabel: Engineering Source
An: 192906513
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
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  Label: Title
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  Data: Synthetic transactivator–promoter systems for exogenous gene expression in Chlamydomonas reinhardtii.
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  Data: <searchLink fieldCode="AR" term="%22Kawabe%2C+Yoshinori%22">Kawabe, Yoshinori</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> kawabe@chem-eng.kyushu-u.ac.jp</i><br /><searchLink fieldCode="AR" term="%22Akiyama%2C+Tatsuki%22">Akiyama, Tatsuki</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Miyazoe%2C+Kokoro%22">Miyazoe, Kokoro</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Kamihira%2C+Masamichi%22">Kamihira, Masamichi</searchLink><relatesTo>1</relatesTo> (AUTHOR)
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Bioscience+%26+Bioengineering%22">Journal of Bioscience & Bioengineering</searchLink>. Jun2026, Vol. 141 Issue 6, p430-437. 8p.
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  Data: <searchLink fieldCode="DE" term="%22Transgene+expression%22">Transgene expression</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+transcription+regulation%22">Genetic transcription regulation</searchLink><br /><searchLink fieldCode="DE" term="%22Transcription+factors%22">Transcription factors</searchLink><br /><searchLink fieldCode="DE" term="%22Protein+expression%22">Protein expression</searchLink><br /><searchLink fieldCode="DE" term="%22Chlamydomonas+reinhardtii%22">Chlamydomonas reinhardtii</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Chlamydomonas reinhardtii , which is a unicellular photosynthetic eukaryote, has long served as a model microalga for fundamental biological research and biotechnological applications. Recently, it has attracted attention as a promising biological resource for the sustainable production of bio-oils and high-value biomolecules. To enhance the biotechnological utility of this species, various genetic engineering tools have been developed in recent years. In this study, the tetracycline repressor (TetR)-based transactivation system, which is widely used in mammalian and other eukaryotic cells, was repurposed to establish a synthetic transcriptional activation system for exogenous gene expression in Chlamydomonas. We first constructed a transient expression-based evaluation platform to screen for transcriptional activation domains (TADs) that are functional in Chlamydomonas. Among the analyzed TADs, VP192 (tandem repeat of 12 copies of the core VP16 domain) had the highest transcriptional activity when fused to either TetR or Gal4 DNA-binding domains. Furthermore, fusion proteins comprising VP192 and either TetR or reverse TetR enabled doxycycline-dependent regulation of transgene expression in a dose-dependent manner. Notably, transcriptional inducibility was maintained even when the tetracycline-responsive element (TRE) was fused to the HSP70A promoter. Combining TetR-VP192 and this synthetic chimeric promoter (TRE-P HSP) yielded transgene expression levels that exceeded those resulting from the strong HSP70A / RbcS2 hybrid promoter by more than 14-fold. These findings suggest that artificial transcription factors and engineered promoters provide a versatile molecular toolkit for regulating exogenous gene expression in Chlamydomonas. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Journal of Bioscience & Bioengineering is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1016/j.jbiosc.2026.02.003
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      – Code: eng
        Text: English
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        PageCount: 8
        StartPage: 430
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      – SubjectFull: Transgene expression
        Type: general
      – SubjectFull: Genetic transcription regulation
        Type: general
      – SubjectFull: Transcription factors
        Type: general
      – SubjectFull: Protein expression
        Type: general
      – SubjectFull: Chlamydomonas reinhardtii
        Type: general
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      – TitleFull: Synthetic transactivator–promoter systems for exogenous gene expression in Chlamydomonas reinhardtii.
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            NameFull: Kawabe, Yoshinori
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            NameFull: Akiyama, Tatsuki
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            NameFull: Miyazoe, Kokoro
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            NameFull: Kamihira, Masamichi
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            – D: 01
              M: 06
              Text: Jun2026
              Type: published
              Y: 2026
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