OPTO-BLUE: An Integrated Bidirectional Optogenetic Lentiviral Platform for Controlled Light-Induced Gene Expression
Keywords: lentivirus, Bidirectional, optogenetic, GAVPO, gene-regulated expression
Abstract
Regulated systems for transgene expression are useful tools in basic research and a promising platform in biomedicine due to their regulated transgene expression by an inducer. The emergence of optogenetics expression systems enabled the construction of light-switchable systems, enhancing the spatial and temporal resolution of a transgene. The LightOn system is an optogenetic tool that regulates the expression of a gene of interest using blue light as an inducer. This system is based on a photosensitive protein (GAVPO), which dimerizes and binds to the UASG sequence in response to blue light, triggering the expression of a downstream transgene. Previously, we adapted the LightOn system to a dual lentiviral vector system for neurons. Here, we continue the optimization and assemble all components of the LightOn system into a single lentiviral plasmid, the OPTO-BLUE system. For functional validation, we used enhanced green fluorescent protein (EGFP) as an expression reporter (OPTO-BLUE-EGFP) and evaluated the efficiency of EGFP expression by transfection and transduction in HEK293-T cells exposed to continuous blue-light illumination. Altogether, these results prove that the optimized OPTO-BLUE system allows the light-controlled expression of a reporter protein according to a specific time and light intensity. Likewise, this system should provide an important molecular tool to modulate gene expression of any protein by blue light.
Más información
Título de la Revista: | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES |
Volumen: | 21 |
Número: | 11 |
Editorial: | MDPI Open Access Publishing |
Fecha de publicación: | 2023 |
Página de inicio: | 9537 |
Página final: | 9547 |
Idioma: | english |
Financiamiento/Sponsor: | NA |
URL: | https://www.mdpi.com/1422-0067/24/11/9537 |
DOI: |
10.3390/ijms24119537 |
Notas: | WOS |