Modulation of Insulin Gene Expression with CRISPR/Cas9-based Transcription Factors

Bakhytzhan Alzhanuly; Zhussipbek Y.  Mukhatayev; Dauren M.  Botbayev; Yeldar  Ashirbekov; Nurlybek D.  Katkenov; Nurlan T.  Dzhaynakbaev; Kamalidin O.  Sharipov

doi:10.3889/oamjms.2021.6980

Authors

Bakhytzhan Alzhanuly Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan; Department of Molecular Biology and Genetics, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, Kazakhstan; Department of Research and Development, Almaty Management University, Almaty, Kazakhstan
Zhussipbek Y. Mukhatayev Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan; Department of Molecular Biology and Genetics, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, Kazakhstan
Dauren M. Botbayev Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan; Department of Molecular Biology and Genetics, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, Kazakhstan
Yeldar Ashirbekov Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan
Nurlybek D. Katkenov Zhangir Khan University, Uralsk, Kazakhstan
Nurlan T. Dzhaynakbaev Kazakh-Russian Medical University, Almaty, Kazakhstan
Kamalidin O. Sharipov Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan

DOI:

https://doi.org/10.3889/oamjms.2021.6980

Keywords:

CRISPR/dCas9 system, Diabetes type I, Embryonic stem cells, HEK293 cells

Abstract

Background: The discovery and use of CRISPR/Cas9 technology have enabled researchers throughout the globe to continuously edit genomes for the benefit of science and medicine. Diabetes type I is one field of medicine where CRISPR/Cas9 has a strong potential for cell therapy development. The long-lasting paucity of healthy cells for clinical transplantation into diabetic patients has led to the search of new methods for producing β-cells from other human cell types. Embryonic stem cells are being studied worldwide as one most promising solution of this need. Aim: The aim of the study is to to check the feasibility of modulating human insulin transcription using CRISPR/Cas9-based synthetic transcription regulation factors.

Results: A new approach for creating potential therapeutic donor cells with enhanced and suppressed insulin production based on one of the latest achievements of human genome editing was developed. Both synthetic transcription activator (VP64) and transcription repressor (KRAB) proteins were shown to function adequately well as a part of the whole CRISPR/Cas9-based system. We claim that our results have a lot to offer and can bring light to many studies where numerous labs are struggling on to treat this disease.

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