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A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique

Genetic screens using CRISPR/Cas9 have been exploited to discover host–virus interactions. These screens have identified viral dependencies on host proteins during their life cycle and potential antiviral strategies. The acyl-CoA binding domain containing 3 (ACBD3) was identified as an essential hos...

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Autores principales: Shin, Hye Jin, Ku, Keun Bon, Kim, Soojin, Kim, Heon Seok, Kim, Yeon-Soo, Kim, Bum-Tae, Kim, Seong-Jun, Kim, Chonsaeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913485/
https://www.ncbi.nlm.nih.gov/pubmed/33546322
http://dx.doi.org/10.3390/v13020237
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author Shin, Hye Jin
Ku, Keun Bon
Kim, Soojin
Kim, Heon Seok
Kim, Yeon-Soo
Kim, Bum-Tae
Kim, Seong-Jun
Kim, Chonsaeng
author_facet Shin, Hye Jin
Ku, Keun Bon
Kim, Soojin
Kim, Heon Seok
Kim, Yeon-Soo
Kim, Bum-Tae
Kim, Seong-Jun
Kim, Chonsaeng
author_sort Shin, Hye Jin
collection PubMed
description Genetic screens using CRISPR/Cas9 have been exploited to discover host–virus interactions. These screens have identified viral dependencies on host proteins during their life cycle and potential antiviral strategies. The acyl-CoA binding domain containing 3 (ACBD3) was identified as an essential host factor for the Coxsackievirus B3 (CVB3) infection. Other groups have also investigated the role of ACBD3 as a host factor for diverse enteroviruses in cultured cells. However, it has not been tested if ACBD3 is required in the animal model of CVB3 infection. Owing to embryonic lethality, conventional knockout mice were not available for in vivo study. As an alternative approach, we used adeno-associated virus (AAV)-mediated CRISPR genome editing to generate mice that lacked ACBD3 within the pancreas, the major target organ for CVB3. Delivery of sgRNAs using self-complementary (sc) AAV8 efficiently induced a loss-of-function mutation in the pancreas of the Cas9 knock-in mice. Loss of ACBD3 in the pancreas resulted in a 100-fold reduction in the CVB3 titer within the pancreas and a noticeable reduction in viral protein expression. These results indicate a crucial function of ACBD3 in CVB3 infection in vivo. AAV-mediated CRISPR genome editing may be applicable to many in vivo studies on the virus–host interaction and identify a novel target for antiviral therapeutics.
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spelling pubmed-79134852021-02-28 A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique Shin, Hye Jin Ku, Keun Bon Kim, Soojin Kim, Heon Seok Kim, Yeon-Soo Kim, Bum-Tae Kim, Seong-Jun Kim, Chonsaeng Viruses Article Genetic screens using CRISPR/Cas9 have been exploited to discover host–virus interactions. These screens have identified viral dependencies on host proteins during their life cycle and potential antiviral strategies. The acyl-CoA binding domain containing 3 (ACBD3) was identified as an essential host factor for the Coxsackievirus B3 (CVB3) infection. Other groups have also investigated the role of ACBD3 as a host factor for diverse enteroviruses in cultured cells. However, it has not been tested if ACBD3 is required in the animal model of CVB3 infection. Owing to embryonic lethality, conventional knockout mice were not available for in vivo study. As an alternative approach, we used adeno-associated virus (AAV)-mediated CRISPR genome editing to generate mice that lacked ACBD3 within the pancreas, the major target organ for CVB3. Delivery of sgRNAs using self-complementary (sc) AAV8 efficiently induced a loss-of-function mutation in the pancreas of the Cas9 knock-in mice. Loss of ACBD3 in the pancreas resulted in a 100-fold reduction in the CVB3 titer within the pancreas and a noticeable reduction in viral protein expression. These results indicate a crucial function of ACBD3 in CVB3 infection in vivo. AAV-mediated CRISPR genome editing may be applicable to many in vivo studies on the virus–host interaction and identify a novel target for antiviral therapeutics. MDPI 2021-02-03 /pmc/articles/PMC7913485/ /pubmed/33546322 http://dx.doi.org/10.3390/v13020237 Text en © 2021 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Shin, Hye Jin
Ku, Keun Bon
Kim, Soojin
Kim, Heon Seok
Kim, Yeon-Soo
Kim, Bum-Tae
Kim, Seong-Jun
Kim, Chonsaeng
A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title_full A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title_fullStr A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title_full_unstemmed A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title_short A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique
title_sort crucial role of acbd3 required for coxsackievirus infection in animal model developed by aav-mediated crispr genome editing technique
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913485/
https://www.ncbi.nlm.nih.gov/pubmed/33546322
http://dx.doi.org/10.3390/v13020237
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