Photoactivatable nanocrispr/cas9 system based on crrna reversibly immobilized on carbon nanoparticles

Olga Semikolenova, Lubov Sakovina, Elizaveta Akhmetova, Daria Kim, Ivan Vokhtantsev, Victor Golyshev, Mariya Vorobyeva, Sergey Novopashin, Darya Novopashina

Результат исследования: Научные публикации в периодических изданияхстатьярецензирование

Аннотация

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.

Язык оригиналаанглийский
Номер статьи10919
ЖурналInternational Journal of Molecular Sciences
Том22
Номер выпуска20
DOI
СостояниеОпубликовано - 1 окт 2021

Предметные области OECD FOS+WOS

  • 1.02 КОМПЬЮТЕРНЫЕ И ИНФОРМАЦИОННЫЕ НАУКИ
  • 2.04 ХИМИЧЕСКИЕ ТЕХНОЛОГИИ
  • 1.04 ХИМИЧЕСКИЕ НАУКИ
  • 1.06 БИОЛОГИЧЕСКИЕ НАУКИ

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