Blue Light-Induced, Dosed Protein Expression of Active BDNF in Human Cells Using the Optogenetic CRY2/CIB System

authored by

Sina Christoffers, Nina Wichert, Elena Wiebe, Maria Leilani Torres-Mapa, Madeleine Goblet, Jennifer Harre, Odett Kaiser, Marc Nils Wahalla, Holger Blume, Alexander Heisterkamp, Athanasia Warnecke, Cornelia Blume

Abstract

The use of optogenetic tools offers an excellent method for spatially and temporally regulated gene and protein expression in cell therapeutic approaches. This could be useful as a concomitant therapeutic measure, especially in small body compartments such as the inner ear, for example, during cochlea implantation, to enhance neuronal cell survival and function. Here, we used the blue light activatable CRY2/CIB system to induce transcription of brain-derived neurotrophic factor (BDNF) in human cells. Transfection with three plasmids, encoding for the optogenetic system and the target, as well as illumination protocols were optimized with luciferase as a reporter to achieve the highest protein expression in human embryonic kidney cells 293. Illumination was performed either with a light-emitting diode or with a scanning laser setup. The optimized protocols were applied for the production of BDNF. We could demonstrate a 64.7-fold increase of BNDF expression upon light induction compared to the basal level. Light-induced BDNF was biologically active and enhanced survival and neurite growth of spiral ganglion neurons. The optogenetic approach can be transferred to autologous cell systems, such as bone marrow-derived mesenchymal stem cells, and thus represents the first optogenetic neurotrophic therapy for the inner ear.

Organisation(s)

Institute of Technical Chemistry
Institute of Microelectronic Systems

External Organisation(s)

Cluster of Excellence Hearing4all
Hannover Medical School (MHH)

Type

Article

Journal

Biotechnology journal

Volume

19

ISSN

1860-6768

Publication date

12.2024

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Applied Microbiology and Biotechnology, Molecular Medicine

Electronic version(s)

https://doi.org/10.1002/biot.202400384 (Access: Open)