Optogenetic control of cancer cell survival in ChR2-transfected HeLa cells

Claudio Córdova, Carlo Lozano, Belén Rodríguez, Ivanny Marchant, Rodrigo Zúñiga, Paola Ochova, Pablo Olivero, Wilfredo Alejandro González-Arriagada

Research output: Contribution to journalArticlepeer-review

Abstract

Optogenetics is a molecular biological technique involving transfection of cells with photosensitive proteins and the subsequent study of their biological effects. The aim of this study was to evaluate the effect of blue light on the survival of HeLa cells, transfected with channelrhodopsin-2 (ChR2). HeLa wild-type cells were transfected with a plasmid that contained the gene for ChR2. Transfection and channel function were evaluated by real-time polymerase chain reaction (RT-PCR), fluorescence imaging using green fluorescent protein (GFP) and flow cytometry for intracellular calcium changes using a Fura Red probe. We developed a platform for optogenetic stimulation for use within the cell culture incubator. Different stimulation procedures using blue light (467 nm) were applied for up to 24 h. Cell survival was determined by flow cytometry using propidium iodide and rhodamine probes. Change in cell survival showed a statistically significant (p < 0.05) inverse association with the frequency and time of application of the light stimulus. This change seemed to be associated with the ChR2 cis-trans-isomerization cycle. Cell death was associated with high concentrations of calcium in the cytoplasm and stimulation intervals less than the period of isomerization. It is possible to transfect HeLa cells with ChR2 and control their survival under blue light stimulation. We suggest that this practice should be considered in the future development of optogenetic systems in biological or biomedical research.

Original languageEnglish
JournalInternational Journal of Experimental Pathology
Early online date17 Nov 2021
DOIs
StateE-pub ahead of print - 17 Nov 2021

Bibliographical note

© 2021 Company of the International Journal of Experimental Pathology (CIJEP).

Keywords

  • genetics
  • light-activated protein
  • physiology
  • rhodopsin
  • transfection

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