CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes

Abstract The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system revolutionized the field of gene editing but viral delivery of the CRISPR/Cas9 system has not been fully explored. Here we adapted clinically relevant high-capacity adenoviral vectors (HCAdV) devoid of all vi...

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Autores principales:	Eric Ehrke-Schulz, Maren Schiwon, Theo Leitner, Stephan Dávid, Thorsten Bergmann, Jing Liu, Anja Ehrhardt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/af66499d641d4b4c9339d899743e75fc
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spelling	oai:doaj.org-article:af66499d641d4b4c9339d899743e75fc2021-12-02T15:05:18ZCRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes10.1038/s41598-017-17180-w2045-2322https://doaj.org/article/af66499d641d4b4c9339d899743e75fc2017-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-17180-whttps://doaj.org/toc/2045-2322Abstract The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system revolutionized the field of gene editing but viral delivery of the CRISPR/Cas9 system has not been fully explored. Here we adapted clinically relevant high-capacity adenoviral vectors (HCAdV) devoid of all viral genes for the delivery of the CRISPR/Cas9 machinery using a single viral vector. We present a platform enabling fast transfer of the Cas9 gene and gRNA expression units into the HCAdV genome including the option to choose between constitutive or inducible Cas9 expression and gRNA multiplexing. Efficacy and versatility of this pipeline was exemplified by producing different CRISPR/Cas9-HCAdV targeting the human papillomavirus (HPV) 18 oncogene E6, the dystrophin gene causing Duchenne muscular dystrophy (DMD) and the HIV co-receptor C-C chemokine receptor type 5 (CCR5). All CRISPR/Cas9-HCAdV proved to be efficient to deliver the respective CRISPR/Cas9 expression units and to introduce the desired DNA double strand breaks at their intended target sites in immortalized and primary cells.Eric Ehrke-SchulzMaren SchiwonTheo LeitnerStephan DávidThorsten BergmannJing LiuAnja EhrhardtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eric Ehrke-Schulz Maren Schiwon Theo Leitner Stephan Dávid Thorsten Bergmann Jing Liu Anja Ehrhardt CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
description	Abstract The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system revolutionized the field of gene editing but viral delivery of the CRISPR/Cas9 system has not been fully explored. Here we adapted clinically relevant high-capacity adenoviral vectors (HCAdV) devoid of all viral genes for the delivery of the CRISPR/Cas9 machinery using a single viral vector. We present a platform enabling fast transfer of the Cas9 gene and gRNA expression units into the HCAdV genome including the option to choose between constitutive or inducible Cas9 expression and gRNA multiplexing. Efficacy and versatility of this pipeline was exemplified by producing different CRISPR/Cas9-HCAdV targeting the human papillomavirus (HPV) 18 oncogene E6, the dystrophin gene causing Duchenne muscular dystrophy (DMD) and the HIV co-receptor C-C chemokine receptor type 5 (CCR5). All CRISPR/Cas9-HCAdV proved to be efficient to deliver the respective CRISPR/Cas9 expression units and to introduce the desired DNA double strand breaks at their intended target sites in immortalized and primary cells.
format	article
author	Eric Ehrke-Schulz Maren Schiwon Theo Leitner Stephan Dávid Thorsten Bergmann Jing Liu Anja Ehrhardt
author_facet	Eric Ehrke-Schulz Maren Schiwon Theo Leitner Stephan Dávid Thorsten Bergmann Jing Liu Anja Ehrhardt
author_sort	Eric Ehrke-Schulz
title	CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
title_short	CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
title_full	CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
title_fullStr	CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
title_full_unstemmed	CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes
title_sort	crispr/cas9 delivery with one single adenoviral vector devoid of all viral genes
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/af66499d641d4b4c9339d899743e75fc
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CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes

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