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Biochemistry (Moscow)

, Volume 84, Issue 3, pp 299–309 | Cite as

A Platform for Studying Neurodegeneration Mechanisms Using Genetically Encoded Biosensors

  • E. I. Ustyantseva
  • S. P. Medvedev
  • A. S. Vetchinova
  • J. M. Minina
  • S. N. Illarioshkin
  • S. M. ZakianEmail author
Article
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Abstract

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.

Keywords

induced pluripotent stem cells biosensors CRISPR/Cas9 

Abbreviations

ALS

amyotrophic lateral sclerosis

CRISPR/Cas9

clustered regularly interspaced short palindromic repeats/CRISPR-associated

EB

embryonic body

ER

endoplasmic reticulum

iPSCs

induced pluripotent stem cells

MBCs

mononuclear blood cells

PBS

phosphate buffered saline

SOD1

superoxide dismutase 1

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Copyright information

© Pleiades Publishing, Ltd. 2019

Authors and Affiliations

  • E. I. Ustyantseva
    • 1
    • 2
    • 3
    • 4
  • S. P. Medvedev
    • 1
    • 2
    • 3
    • 4
  • A. S. Vetchinova
    • 5
  • J. M. Minina
    • 1
  • S. N. Illarioshkin
    • 5
  • S. M. Zakian
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Federal Research Center Institute of Cytology and GeneticsSiberian Branch of the Russian Academy of SciencesNovosibirskRussia
  2. 2.Institute of Chemical Biology and Fundamental MedicineSiberian Branch of the Russian Academy of SciencesNovosibirskRussia
  3. 3.Meshalkin National Medical Research CenterMinistry of Health of the Russian FederationNovosibirskRussia
  4. 4.Novosibirsk State UniversityNovosibirskRussia
  5. 5.Research Center of NeurologyMoscowRussia

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