A Platform for Studying Neurodegeneration Mechanisms Using Genetically Encoded Biosensors

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E. I. Ustyantseva^1,2,3,4, S. P. Medvedev^1,2,3,4, A. S. Vetchinova⁵, J. M. Minina¹, S. N. Illarioshkin⁵, and S. M. Zakian^1,2,3,4,a*

¹Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia

²Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia

³Meshalkin National Medical Research Center, Ministry of Health of the Russian Federation, 630055 Novosibirsk, Russia

⁴Novosibirsk State University, 630090 Novosibirsk, Russia

⁵Research Center of Neurology, 125367 Moscow, Russia

^* To whom correspondence should be addressed.

Received October 14, 2018; Revised November 11, 2018; Accepted November 11, 2018

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Patient-specific induced pluripotent stem cells (iPSCs) capable of differentiation into required cell type are a promising model for studying various pathological processes and development of new therapeutic approaches. However, no conventional strategies for using iPSCs in disease research have been established yet. Genetically encoded biosensors can be used for monitoring messenger molecules, metabolites, and enzyme activity in real time with the following conversion of the registered signals in quantitative data, thus allowing evaluation of the impact of certain molecules on pathology development. In this article, we describe the development of a universal cell-based platform for studying pathological processes associated with amyotrophic lateral sclerosis. For this purpose, we have created a series of plasmid constructs for monitoring endoplasmic reticulum stress, oxidative stress, apoptosis, and Ca²⁺-dependent hyperexcitability and generated transgenic iPSC line carrying mutation in the superoxide dismutase 1 gene (SOD1) and healthy control cell line. Both cell lines have specific transactivator sequence required for doxycycline-controlled transcriptional activation and can be used for a single-step biosensor insertion.

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KEY WORDS: induced pluripotent stem cells, biosensors, CRISPR/Cas9

DOI: 10.1134/S000629791903012X

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