Photoactivatable CRISPR/Cas9 System

Photoactivatable CRISPR/Cas9 System

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed. Photocleavable oligodeoxyribonucleotides contain two or three photodegradable linkers in their stru...

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Veröffentlicht in:Russian journal of bioorganic chemistry 2021-03, Vol.47 (2), p.496-504
Hauptverfasser: Akhmetova, E. A., Golyshev, V. M., Vokhtantcev, I. P., Meschaninova, M. I., Venyaminova, A. G., Novopashina, D. S.
Format: Artikel
Sprache:eng
Schlagworte:
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
CRISPR
Genetic modification
Life Sciences
Oligonucleotides
Organic Chemistry
Structural stability
Thermal stability
Ultraviolet radiation
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Zusammenfassung:A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed. Photocleavable oligodeoxyribonucleotides contain two or three photodegradable linkers in their structure. The thermal stability of complexes of these modified oligonucleotides with guide crRNA decreases with increasing the number of linkers. The kinetics of the photomodified oligodeoxyribonucleotides cleavage has been investigated. It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
ISSN:1068-1620
1608-330X
DOI:10.1134/S1068162021020023