Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor

A novel Cas13d-based gene-editing approach has been developed to target viral RNAs in infected cells, reducing the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Zika virus (ZIKV) by up to 90% compared with controls. Despite its potential, the use of Cas13d as an ant...

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Main Authors: Alessandro De Carli, Domenico Favaro, Carolina Filipponi, Fabio Filippini, Rossella Fonnesu, Erika Plicanti, Silvia Nottoli, Piotr Barski, Agnieszka Lindstaedt, Dariusz Witt, Alessandra Falleni, Giada Frenzilli, Ana Alcalá-Lalinde, Elena Herrera-Carrillo, Vittoria Raffa, Giulia Freer, Mauro Pistello, Michele Lai
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Molecular Therapy: Nucleic Acids
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Online Access:http://www.sciencedirect.com/science/article/pii/S2162253125000940
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author Alessandro De Carli
Domenico Favaro
Carolina Filipponi
Fabio Filippini
Rossella Fonnesu
Erika Plicanti
Silvia Nottoli
Piotr Barski
Agnieszka Lindstaedt
Dariusz Witt
Alessandra Falleni
Giada Frenzilli
Ana Alcalá-Lalinde
Elena Herrera-Carrillo
Vittoria Raffa
Giulia Freer
Mauro Pistello
Michele Lai
author_facet Alessandro De Carli
Domenico Favaro
Carolina Filipponi
Fabio Filippini
Rossella Fonnesu
Erika Plicanti
Silvia Nottoli
Piotr Barski
Agnieszka Lindstaedt
Dariusz Witt
Alessandra Falleni
Giada Frenzilli
Ana Alcalá-Lalinde
Elena Herrera-Carrillo
Vittoria Raffa
Giulia Freer
Mauro Pistello
Michele Lai
author_sort Alessandro De Carli
collection DOAJ
description A novel Cas13d-based gene-editing approach has been developed to target viral RNAs in infected cells, reducing the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Zika virus (ZIKV) by up to 90% compared with controls. Despite its potential, the use of Cas13d as an antiviral faces several challenges that limit its effectiveness before reaching target cells. This study presents a proof-of-concept strategy for constructing Cas13d with gold nanoparticles (Au_NPs) to destroy SARS-CoV-2 and ZIKV genomes into cells. The Au_NPs Cas13d complexes were administered to Huh-7 cells infected with either virus, in single or multiple doses. The study demonstrated that Au_NPs Cas13d cuts target RNAs with comparable efficiency as lipofected ribonucleoprotein (RNP). Additionally, we found that Au_NPs Cas13d can spontaneously enter cells by endocytosis or diffusion, before the first 4 h of treatment. Au_NPs Cas13d co-localized with SARS-CoV-2 virions in early endosomes and reduced SARS-CoV-2 replication after a single administration, unlike RNPs, which showed no antiviral activity. However, Au_NPs Cas13d was less efficient at reducing ZIKV replication compared with lipofected Cas13d-RNPs, likely due to different intracellular localization. These results suggest that Au_NPs can be adapted as a new antiviral strategy, highlighting an innovative delivery method of Cas13d against viruses without the need for transfecting, providing a new gene-editing-based approach against emerging RNA viruses.
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spelling doaj-art-cb60d0819c74446f8222f8d08da720cb2025-08-20T02:13:23ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-06-0136210254010.1016/j.omtn.2025.102540Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armorAlessandro De Carli0Domenico Favaro1Carolina Filipponi2Fabio Filippini3Rossella Fonnesu4Erika Plicanti5Silvia Nottoli6Piotr Barski7Agnieszka Lindstaedt8Dariusz Witt9Alessandra Falleni10Giada Frenzilli11Ana Alcalá-Lalinde12Elena Herrera-Carrillo13Vittoria Raffa14Giulia Freer15Mauro Pistello16Michele Lai17Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy; Retrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy; Centre for Instrumentation Sharing, University of Pisa (CISUP), 56100 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy; Virology Unit, Pisa University Hospital, 56124 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, ItalyVirology Unit, Pisa University Hospital, 56124 Pisa, ItalyDepartment of Medical Biotechnologies, University of Siena, 53100 Siena, Italy; Retrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, ItalyVirology Unit, Pisa University Hospital, 56124 Pisa, ItalyProChimia Surfaces Sp. z o.o., Al Zwycięstwa 96/98 F8, 81-451 Gdynia, PolandProChimia Surfaces Sp. z o.o., Al Zwycięstwa 96/98 F8, 81-451 Gdynia, PolandProChimia Surfaces Sp. z o.o., Al Zwycięstwa 96/98 F8, 81-451 Gdynia, PolandDepartment of Clinical and Experimental Medicine, Section of Applied Biology and Genetics, and INSTM Local Unit, University of Pisa, 56126 Pisa, ItalyDepartment of Clinical and Experimental Medicine, Section of Applied Biology and Genetics, and INSTM Local Unit, University of Pisa, 56126 Pisa, ItalyAmsterdam UMC, University of Amsterdam, Medical Microbiology and Infection Prevention, Meibergdreef 9, Amsterdam 1105AZ, the Netherlands; Amsterdam Institute for Immunology and Infectious Diseases, Amsterdam 1105AZ, the NetherlandsAmsterdam UMC, University of Amsterdam, Medical Microbiology and Infection Prevention, Meibergdreef 9, Amsterdam 1105AZ, the Netherlands; Amsterdam Institute for Immunology and Infectious Diseases, Amsterdam 1105AZ, the NetherlandsDepartment of Biology, Università di Pisa, 56127 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy; Centre for Instrumentation Sharing, University of Pisa (CISUP), 56100 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy; Virology Unit, Pisa University Hospital, 56124 Pisa, ItalyRetrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy; Centre for Instrumentation Sharing, University of Pisa (CISUP), 56100 Pisa, Italy; Corresponding author: Michele Lai, Retrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy.A novel Cas13d-based gene-editing approach has been developed to target viral RNAs in infected cells, reducing the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Zika virus (ZIKV) by up to 90% compared with controls. Despite its potential, the use of Cas13d as an antiviral faces several challenges that limit its effectiveness before reaching target cells. This study presents a proof-of-concept strategy for constructing Cas13d with gold nanoparticles (Au_NPs) to destroy SARS-CoV-2 and ZIKV genomes into cells. The Au_NPs Cas13d complexes were administered to Huh-7 cells infected with either virus, in single or multiple doses. The study demonstrated that Au_NPs Cas13d cuts target RNAs with comparable efficiency as lipofected ribonucleoprotein (RNP). Additionally, we found that Au_NPs Cas13d can spontaneously enter cells by endocytosis or diffusion, before the first 4 h of treatment. Au_NPs Cas13d co-localized with SARS-CoV-2 virions in early endosomes and reduced SARS-CoV-2 replication after a single administration, unlike RNPs, which showed no antiviral activity. However, Au_NPs Cas13d was less efficient at reducing ZIKV replication compared with lipofected Cas13d-RNPs, likely due to different intracellular localization. These results suggest that Au_NPs can be adapted as a new antiviral strategy, highlighting an innovative delivery method of Cas13d against viruses without the need for transfecting, providing a new gene-editing-based approach against emerging RNA viruses.http://www.sciencedirect.com/science/article/pii/S2162253125000940MT: RNA/DNA EditingSARS-CoV-2Zika virusCas13dgold nanoparticlesAu_NPs Cas13d
spellingShingle Alessandro De Carli
Domenico Favaro
Carolina Filipponi
Fabio Filippini
Rossella Fonnesu
Erika Plicanti
Silvia Nottoli
Piotr Barski
Agnieszka Lindstaedt
Dariusz Witt
Alessandra Falleni
Giada Frenzilli
Ana Alcalá-Lalinde
Elena Herrera-Carrillo
Vittoria Raffa
Giulia Freer
Mauro Pistello
Michele Lai
Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
Molecular Therapy: Nucleic Acids
MT: RNA/DNA Editing
SARS-CoV-2
Zika virus
Cas13d
gold nanoparticles
Au_NPs Cas13d
title Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
title_full Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
title_fullStr Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
title_full_unstemmed Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
title_short Fighting RNA viruses with a gold nanoparticle Cas13d gene-editing armor
title_sort fighting rna viruses with a gold nanoparticle cas13d gene editing armor
topic MT: RNA/DNA Editing
SARS-CoV-2
Zika virus
Cas13d
gold nanoparticles
Au_NPs Cas13d
url http://www.sciencedirect.com/science/article/pii/S2162253125000940
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