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Optimization of the Light-On system in a lentiviral platform to a light-controlled expression of genes in neurons
Zárate, Rafaella V; Arancibia, Duxan; Fernández, Anllely; Signorelli, Janetti R; F. Larrondo, Luis; Andrés, María Estela; Zamorano, Pedro.
  • Zárate, Rafaella V; Universidad de Antofagasta. Facultad de Ciencias de la Salud. Departamento Biomédico. Antofagasta. CL
  • Arancibia, Duxan; Universidad de Antofagasta. Facultad de Ciencias de la Salud. Departamento Biomédico. Antofagasta. CL
  • Fernández, Anllely; Universidad de Antofagasta. Facultad de Ciencias de la Salud. Departamento Biomédico. Antofagasta. CL
  • Signorelli, Janetti R; Universidad de Antofagasta. Facultad de Ciencias de la Salud. Departamento Biomédico. Antofagasta. CL
  • F. Larrondo, Luis; Pontificia Universidad Católica de Chile. Facultad de Ciencias Biológicas. Departamento de Genética Molecular y Microbiología. Santiago. CL
  • Andrés, María Estela; Pontificia Universidad Católica de Chile. Facultad de Ciencias Biológicas. Departamento de Biología Celular y Molecular. Santiago. CL
  • Zamorano, Pedro; Universidad de Antofagasta. Facultad de Ciencias de la Salud. Departamento Biomédico. Antofagasta. CL
Electron. j. biotechnol ; 51: 50-57, May. 2021. ilus, graf
Article in English | LILACS | ID: biblio-1343384
ABSTRACT

BACKGROUND:

Molecular brain therapies require the development of molecular switches to control gene expression in a limited and regulated manner in time and space. Light-switchable gene systems allow precise control of gene expression with an enhanced spatio-temporal resolution compared to chemical inducers. In this work, we adapted the existing light-switchable Light-On system into a lentiviral platform, which consists of two modules (i) one for the expression of the blue light-switchable transactivator GAVPO and (ii) a second module containing an inducible-UAS promoter (UAS) modulated by a light-activated GAVPO.

RESULTS:

In the HEK293-T cell line transfected with this lentiviral plasmids system, the expression of the reporter mCherry increased between 4 to 5 fold after light induction. A time expression analysis after light induction during 24 h revealed that mRNA levels continuously increased up to 9 h, while protein levels increased throughout the experiment. Finally, transduction of cultured rat hippocampal neurons with this dual Light-On lentiviral system showed that CDNF, a potential therapeutic trophic factor, was induced only in cells exposed to blue light.

CONCLUSIONS:

In conclusion, the optimized lentiviral platform of the Light-On system provides an efficient way to control gene expression in neurons, suggesting that this platform could potentially be used in biomedical and neuroscience research, and eventually in brain therapies for neurodegenerative diseases.
Subject(s)

Full text: Available Index: LILACS (Americas) Main subject: Gene Expression Regulation / Optogenetics / Light / Neurons Language: English Journal: Electron. j. biotechnol Journal subject: Biotechnology Year: 2021 Type: Article Affiliation country: Chile Institution/Affiliation country: Pontificia Universidad Católica de Chile/CL / Universidad de Antofagasta/CL

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Full text: Available Index: LILACS (Americas) Main subject: Gene Expression Regulation / Optogenetics / Light / Neurons Language: English Journal: Electron. j. biotechnol Journal subject: Biotechnology Year: 2021 Type: Article Affiliation country: Chile Institution/Affiliation country: Pontificia Universidad Católica de Chile/CL / Universidad de Antofagasta/CL