Research output: Contribution to journal › Article › peer-review
Photoactivatable nanocrispr/cas9 system based on crrna reversibly immobilized on carbon nanoparticles. / Semikolenova, Olga; Sakovina, Lubov; Akhmetova, Elizaveta et al.
In: International Journal of Molecular Sciences, Vol. 22, No. 20, 10919, 01.10.2021.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Photoactivatable nanocrispr/cas9 system based on crrna reversibly immobilized on carbon nanoparticles
AU - Semikolenova, Olga
AU - Sakovina, Lubov
AU - Akhmetova, Elizaveta
AU - Kim, Daria
AU - Vokhtantsev, Ivan
AU - Golyshev, Victor
AU - Vorobyeva, Mariya
AU - Novopashin, Sergey
AU - Novopashina, Darya
N1 - Funding Information: Funding: The reported study was funded by RFBR, project number 19-34-51026, and the Russian State funded budget project of ICBFM SB RAS N 121031300042-1. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/10/1
Y1 - 2021/10/1
N2 - 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.
AB - 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.
KW - Carbon nanoparti-cles
KW - CRISPR/Cas9 gene editing system
KW - CrRNA immobilization
KW - Magnetic nanoparticles
KW - Photoactivation
KW - Photocleavable oligonucleotides
KW - Pyrene
UR - http://www.scopus.com/inward/record.url?scp=85116735112&partnerID=8YFLogxK
U2 - 10.3390/ijms222010919
DO - 10.3390/ijms222010919
M3 - Article
C2 - 34681578
AN - SCOPUS:85116735112
VL - 22
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
SN - 1661-6596
IS - 20
M1 - 10919
ER -
ID: 34410283