Near-Infrared Light-Controlled Gene Expression and Protein Targeting in Neurons and Non-neuronal Cells.
Taras A RedchukMaksim M KarasevEvgeniya S OmelinaVladislav V VerkhushaPublished in: Chembiochem : a European journal of chemical biology (2018)
Near-infrared (NIR) light-inducible binding of bacterial phytochrome BphP1 to its engineered partner, QPAS1, is used for optical protein regulation in mammalian cells. However, there are no data on the application of the BphP1-QPAS1 pair in cells derived from various mammalian tissues. Here, we tested the functionality of two BphP1-QPAS1-based optogenetic tools-an NIR- and blue-light-sensing system for control of protein localization (iRIS) and an NIR light-sensing system for transcription activation (TA)-in several cell types, including cortical neurons. We found that the performance of these optogenetic tools often relied on physiological properties of a specific cell type, such as nuclear transport, which could limit the applicability of the blue-light-sensitive component of iRIS. In contrast, the NIR-light-sensing component of iRIS performed well in all tested cell types. The TA system showed the best performance in cervical cancer (HeLa), bone cancer (U-2 OS), and human embryonic kidney (HEK-293) cells. The small size of the QPAS1 component allowed the design of adeno-associated virus (AAV) particles, which were applied to deliver the TA system to neurons.
Keyphrases
- induced apoptosis
- gene expression
- cell cycle arrest
- photodynamic therapy
- spinal cord
- fluorescent probe
- drug release
- single cell
- endothelial cells
- high resolution
- stem cells
- binding protein
- dna methylation
- young adults
- squamous cell carcinoma
- cell proliferation
- transcription factor
- mesenchymal stem cells
- bone marrow
- body composition
- papillary thyroid
- contrast enhanced
- men who have sex with men
- soft tissue
- hiv infected
- hiv testing
- bone regeneration