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The Efficiency of Gene Activation Using CRISPR/dCas9-Based Transactivation Systems Depends on the System Run Time

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Abstract

Transactivation systems are a promising application based on the CRISPR/Cas9 system and allow targeted control of gene expression levels in cell culture. However, their performance has been reported to vary considerably depending on the cell type and the activator system. Three activator systems (dCas9-VP160, dCas9-SunTag, and dCas9-VPR) were compared for the efficiency of activating expression of OCT4, NANOG, PDX1, FOXA2, NKX2-2, and NKX6-1 in an immortalized human skin fibroblast line. The activation efficiency was found to depend on the activation system type; the extent of activation depended on the system run time.

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Funding

This work was supported by the Ministry of Science and Higher Education (project no. 075-15-2019-1789, Center of Precise Genome Editing and Genetic Technologies for Biomedicine). Studies by immunofluorescence microscopy were supported by the Russian Science Foundation (project no. 17-75-20178). A.S. Artyuhov acknowledges a Systems Biology Fellowship grant from the Skolkovo Institute of Science and Technology.

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Authors and Affiliations

  1. Center of Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997, Moscow, Russia

    A. S. Artyuhov, A. V. Sorokina, K. M. Shakirova, E. D. Momotyuk & E. B. Dashinimaev

  2. Moscow Institute of Physics and Technology (State University), 141701, Dolgoprudny, Moscow oblast, Russia

    D. A. Dorovskiy & E. B. Dashinimaev

  3. Koltsov Institute of Developmental Biology, Russian Academy of Sciences, 119334, Moscow, Russia

    E. D. Momotyuk

Authors
  1. A. S. Artyuhov
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  2. D. A. Dorovskiy
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  3. A. V. Sorokina
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  4. K. M. Shakirova
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  5. E. D. Momotyuk
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  6. E. B. Dashinimaev
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Correspondence to E. B. Dashinimaev.

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The authors declare that they have no conflicts of interest. This article does not contain any studies involving animals or human subjects performed by any of the authors.

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Translated by T. Tkacheva

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Artyuhov, A.S., Dorovskiy, D.A., Sorokina, A.V. et al. The Efficiency of Gene Activation Using CRISPR/dCas9-Based Transactivation Systems Depends on the System Run Time. Mol Biol 56, 942–949 (2022). https://doi.org/10.1134/S0026893322060048

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