Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles

Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles Full article

Journal

International Journal of Molecular Sciences
, E-ISSN: 1422-0067

Output data

Year: 2021, Volume: 22, Number: 20, Article number : 10919, Pages count : DOI: 10.3390/ijms222010919

Tags

CRISPR/Cas9 gene editing system; photocleavable oligonucleotides; carbon nanoparticles; magnetic nanoparticles; crRNA immobilization; pyrene; photoactivation

Authors

Semikolenova Olga ^1,2 , Sakovina Lubov ^1,2 , Akhmetova Elizaveta ³ , Kim Daria ^1,2 , Vokhtantsev Ivan ^1,3 , Golyshev Victor ¹ , Vorobyeva Mariya ¹ , Novopashin Sergey ⁴ , Novopashina Darya ^1,2,3

Affiliations

1	(Данные Web of science) Inst Chem Biol & Fundamental Med SB RAS, Novosibirsk 630090, Russia
2	(Данные Web of science) Scientific Center of Genetic and Life Sciences, Sirius University of Science and Technology, 354000 Sochi, Russia
3	(Данные Web of science) Novosibirsk State Univ, Novosibirsk 630090, Russia
4	(Данные Web of science) Kutateladze Inst Thermophys SB RAS, Novosibirsk 630090, Russia

Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA. PC-DNAs contained up to three UV-sensitive linkers made of 1-(2-nitrophenyl)-1,2-ethanediol inside the oligonucleotide chain. Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3 '-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity. We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.

Semikolenova O. , Sakovina L. , Akhmetova E. , Kim D. , Vokhtantsev I. , Golyshev V. , Vorobyeva M. , Novopashin S. , Novopashina D.
Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles
International Journal of Molecular Sciences. 2021. V.22. N20. 10919 . DOI: 10.3390/ijms222010919 WOS Scopus РИНЦ OpenAlex

Published print:

Oct 9, 2021

Web of science:	WOS:000717177700001
Scopus:	2-s2.0-85116735112
Elibrary:	47511880
OpenAlex:	W3205498202

DB	Citing
Web of science	3
Scopus	2
Elibrary	4
OpenAlex	4