Photoactivatable CRISPR/Cas9 System

E. A. Akhmetova; V. M. Golyshev; I. P. Vokhtantcev; M. I. Meschaninova; Aliya G. Venyaminova; D. S. Novopashina

doi:10.1134/S1068162021020023

Photoactivatable CRISPR/Cas9 System

E. A. Akhmetova, V. M. Golyshev, I. P. Vokhtantcev, M. I. Meschaninova, Aliya G. Venyaminova, D. S. Novopashina

Результат исследования: Научные публикации в периодических изданиях › статья › рецензирование

Аннотация

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.

Язык оригинала	английский
Страницы (с-по)	496-504
Число страниц	9
Журнал	Russian Journal of Bioorganic Chemistry
Том	47
Номер выпуска	2
DOI	https://doi.org/10.1134/S1068162021020023
Состояние	Опубликовано - мар. 2021

Предметные области OECD FOS+WOS

1.06 БИОЛОГИЧЕСКИЕ НАУКИ
1.04 ХИМИЧЕСКИЕ НАУКИ
1.06.CQ БИОХИМИЯ И МОЛЕКУЛЯРНАЯ БИОЛОГИЯ

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10.1134/S1068162021020023

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title = "Photoactivatable CRISPR/Cas9 System",

abstract = "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.",

keywords = "guide RNAs for CRISPR/Cas9 system, photoactivatable gene editing in vitro, photoblocking oligodeoxyribonucleotide, photocleavable linker",

author = "Akhmetova, {E. A.} and Golyshev, {V. M.} and Vokhtantcev, {I. P.} and Meschaninova, {M. I.} and Venyaminova, {Aliya G.} and Novopashina, {D. S.}",

note = "Funding Information: The work was supported by the Russian Foundation of Basic Research (grant no. 19-34-51026). Optimization of the synthesis of extended oligodeoxyribonucleotides was supported by State-funded budget projects (Program of fundamental scientific research of the State Academies of Sciences for 2013–2020 [АААА-А17-117020210021-7]). Publisher Copyright: {\textcopyright} 2021, Pleiades Publishing, Ltd. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = mar,

doi = "10.1134/S1068162021020023",

language = "English",

volume = "47",

pages = "496--504",

journal = "Russian Journal of Bioorganic Chemistry",

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AU - Akhmetova, E. A.

AU - Golyshev, V. M.

AU - Vokhtantcev, I. P.

AU - Meschaninova, M. I.

AU - Venyaminova, Aliya G.

AU - Novopashina, D. S.

N1 - Funding Information: The work was supported by the Russian Foundation of Basic Research (grant no. 19-34-51026). Optimization of the synthesis of extended oligodeoxyribonucleotides was supported by State-funded budget projects (Program of fundamental scientific research of the State Academies of Sciences for 2013–2020 [АААА-А17-117020210021-7]). Publisher Copyright: © 2021, Pleiades Publishing, Ltd. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/3

Y1 - 2021/3

N2 - 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.

AB - 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.

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Photoactivatable CRISPR/Cas9 System

Аннотация

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