Cargando…
Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles
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...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8539621/ https://www.ncbi.nlm.nih.gov/pubmed/34681578 http://dx.doi.org/10.3390/ijms222010919 |
_version_ | 1784588791366811648 |
---|---|
author | Semikolenova, Olga Sakovina, Lubov Akhmetova, Elizaveta Kim, Daria Vokhtantsev, Ivan Golyshev, Victor Vorobyeva, Mariya Novopashin, Sergey Novopashina, Darya |
author_facet | Semikolenova, Olga Sakovina, Lubov Akhmetova, Elizaveta Kim, Daria Vokhtantsev, Ivan Golyshev, Victor Vorobyeva, Mariya Novopashin, Sergey Novopashina, Darya |
author_sort | Semikolenova, Olga |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8539621 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-85396212021-10-24 Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles Semikolenova, Olga Sakovina, Lubov Akhmetova, Elizaveta Kim, Daria Vokhtantsev, Ivan Golyshev, Victor Vorobyeva, Mariya Novopashin, Sergey Novopashina, Darya Int J Mol Sci Article 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. MDPI 2021-10-09 /pmc/articles/PMC8539621/ /pubmed/34681578 http://dx.doi.org/10.3390/ijms222010919 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Semikolenova, Olga Sakovina, Lubov Akhmetova, Elizaveta Kim, Daria Vokhtantsev, Ivan Golyshev, Victor Vorobyeva, Mariya Novopashin, Sergey Novopashina, Darya Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title | Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title_full | Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title_fullStr | Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title_full_unstemmed | Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title_short | Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles |
title_sort | photoactivatable nanocrispr/cas9 system based on crrna reversibly immobilized on carbon nanoparticles |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8539621/ https://www.ncbi.nlm.nih.gov/pubmed/34681578 http://dx.doi.org/10.3390/ijms222010919 |
work_keys_str_mv | AT semikolenovaolga photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT sakovinalubov photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT akhmetovaelizaveta photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT kimdaria photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT vokhtantsevivan photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT golyshevvictor photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT vorobyevamariya photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT novopashinsergey photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles AT novopashinadarya photoactivatablenanocrisprcas9systembasedoncrrnareversiblyimmobilizedoncarbonnanoparticles |