Details
| Original language | English |
|---|---|
| Article number | e202400384 |
| Journal | Biotechnology journal |
| Volume | 19 |
| Issue number | 12 |
| Publication status | Published - Dec 2024 |
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.
Keywords
- BDNF, cochlea implant, gene expression, HEK293, neurotrophins, spiral ganglion neurons
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
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In: Biotechnology journal, Vol. 19, No. 12, e202400384, 12.2024.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Blue Light-Induced, Dosed Protein Expression of Active BDNF in Human Cells Using the Optogenetic CRY2/CIB System
AU - Christoffers, Sina
AU - Wichert, Nina
AU - Wiebe, Elena
AU - Torres-Mapa, Maria Leilani
AU - Goblet, Madeleine
AU - Harre, Jennifer
AU - Kaiser, Odett
AU - Wahalla, Marc Nils
AU - Blume, Holger
AU - Heisterkamp, Alexander
AU - Warnecke, Athanasia
AU - Blume, Cornelia
N1 - Publisher Copyright: © 2024 The Author(s). Biotechnology Journal published by Wiley-VCH GmbH.
PY - 2024/12
Y1 - 2024/12
N2 - 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.
AB - 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.
KW - BDNF
KW - cochlea implant
KW - gene expression
KW - HEK293
KW - neurotrophins
KW - spiral ganglion neurons
UR - http://www.scopus.com/inward/record.url?scp=85213083651&partnerID=8YFLogxK
U2 - 10.1002/biot.202400384
DO - 10.1002/biot.202400384
M3 - Article
AN - SCOPUS:85213083651
VL - 19
JO - Biotechnology journal
JF - Biotechnology journal
SN - 1860-6768
IS - 12
M1 - e202400384
ER -