MDPI - Publisher of Open Access Journals

15 pages, 1936 KiB

Open AccessArticle

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

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

Int. J. Mol. Sci. 2021, 22(20), 10919; https://doi.org/10.3390/ijms222010919 - 9 Oct 2021

Cited by 10 | Viewed by 2742

Abstract

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. Full article

(This article belongs to the Special Issue Interactions of Nanoparticles with Biomolecules)

► Show Figures

Figure 1

11 pages, 1319 KiB

Open AccessArticle

Practical Synthesis of Quinoline-Protected Morpholino Oligomers for Light-Triggered Regulation of Gene Function

by Davide Deodato and Timothy M. Dore

Molecules 2020, 25(9), 2078; https://doi.org/10.3390/molecules25092078 - 29 Apr 2020

Cited by 15 | Viewed by 8666

Abstract

Photoactivatable cyclic caged morpholino oligomers (ccMOs) represent a promising tool to selectively regulate gene expression with spatiotemporal control. Nevertheless, some challenges associated with the preparation of these reagents have limited their broader use in biological settings. We describe a novel ccMO design that overcomes many of the challenges and considerably expedites the synthetic preparation. The key factor is the introduction of an ethynyl function on the photocleavable linker to facilitate the use of a Huisgen 1,3-dipolar cycloaddition for the coupling reaction with the oligonucleotide. Compared to previous strategies, this modification reduces the number of synthetic steps and significantly improves the total yield and the stability of the linker. We used the alkynyl-functionalized linker for the preparation of two different ccMOs targeting the mRNA of the glutamic acid decarboxylase genes, gad1 and gad2. HPLC analysis confirms that the caging strategy successfully inhibits the DNA binding ability, and the activity can be restored by brief illumination with 405-nm light. Overall, the straightforward preparation together with the clean and fast photochemistry make these caged antisense reagents excellent tools to modulate gene function in-vivo with spatial and temporal precision. Full article

(This article belongs to the Special Issue Synthetic Heterocyclic Chemistry)

► Show Figures

Graphical abstract

12 pages, 875 KiB

Open AccessArticle

Use of Oligonucleotides Carrying Photolabile Groups for the Control of the Deposition of Nanoparticles in Surfaces and Nanoparticle Association

by Brendan Manning and Ramon Eritja

Int. J. Mol. Sci. 2011, 12(10), 7238-7249; https://doi.org/10.3390/ijms12107238 - 24 Oct 2011

Cited by 3 | Viewed by 6824

Abstract

An oligodeoxynucleotide hairpin containing a photolabile 2-nitrobenzyl group in the loop and terminated with a thiol function was prepared. The photocleavage of such a hairpin on gold yields a surface activated with a single stranded oligonucleotide which can be utilised to direct the assembly of nanoparticles conjugated with a complementary strand. Analysis of photocleaved surfaces gives nanoparticle coverage one order of magnitude higher than nonphotocleaved surfaces. This illustrates the ability of photocleavable hairpins to direct the assembly of nanomaterials on conducting materials. The conjugation of the photocleavable hairpin to a gold nanoparticle allows the observation of intermolecular interactions between hairpins linked in different nanoparticles, by comparing the thermal dissociations of a hairpin-nanoparticle conjugates at 260 nm and 520 nm. We have also shown that it is possible to permanently alter the physiochemical properties of DNA-nanoparticles by the introduction of a photocleavable group. Indeed for the first time it has been shown that by exposure to UV light the disassembly of nanoparticle aggregates can be induced. Full article

(This article belongs to the Section Materials Science)

► Show Figures

Graphical abstract

Search Results (3)

Further Information

Guidelines

MDPI Initiatives

Follow MDPI

Saved Queries

Search Filter Reset All

Years

Feature Papers

Subjects

Journals

Article Types

Countries / Regions

Search Results (3)

Further Information

Guidelines

MDPI Initiatives

Follow MDPI