A novel cloning strategy for one-step assembly of multiplex CRISPR vectors

One key advantage of the CRISPR/Cas9 system in comparison with other gene editing approaches lies in its potential for multiplexing. Here, we describe an elaborate procedure that allows the assembly of multiple gRNA expression cassettes into a vector of choice within a single step, termed ASAP(Adapt...

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Autores principales: Zuckermann, Marc, Hlevnjak, Mario, Yazdanparast, Haniyeh, Zapatka, Marc, Jones, David T. W., Lichter, Peter, Gronych, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6269432/
https://www.ncbi.nlm.nih.gov/pubmed/30504793
http://dx.doi.org/10.1038/s41598-018-35727-3
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author Zuckermann, Marc
Hlevnjak, Mario
Yazdanparast, Haniyeh
Zapatka, Marc
Jones, David T. W.
Lichter, Peter
Gronych, Jan
author_facet Zuckermann, Marc
Hlevnjak, Mario
Yazdanparast, Haniyeh
Zapatka, Marc
Jones, David T. W.
Lichter, Peter
Gronych, Jan
author_sort Zuckermann, Marc
collection PubMed
description One key advantage of the CRISPR/Cas9 system in comparison with other gene editing approaches lies in its potential for multiplexing. Here, we describe an elaborate procedure that allows the assembly of multiple gRNA expression cassettes into a vector of choice within a single step, termed ASAP(Adaptable System for Assembly of multiplexed Plasmids)-cloning. We demonstrate the utility of ASAP-cloning for multiple CRISPR-mediated applications, including efficient multiplex gene editing, robust transcription activation and convenient analysis of Cas9 activity in the presence of multiple gRNAs.
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spelling pubmed-62694322018-12-04 A novel cloning strategy for one-step assembly of multiplex CRISPR vectors Zuckermann, Marc Hlevnjak, Mario Yazdanparast, Haniyeh Zapatka, Marc Jones, David T. W. Lichter, Peter Gronych, Jan Sci Rep Article One key advantage of the CRISPR/Cas9 system in comparison with other gene editing approaches lies in its potential for multiplexing. Here, we describe an elaborate procedure that allows the assembly of multiple gRNA expression cassettes into a vector of choice within a single step, termed ASAP(Adaptable System for Assembly of multiplexed Plasmids)-cloning. We demonstrate the utility of ASAP-cloning for multiple CRISPR-mediated applications, including efficient multiplex gene editing, robust transcription activation and convenient analysis of Cas9 activity in the presence of multiple gRNAs. Nature Publishing Group UK 2018-11-30 /pmc/articles/PMC6269432/ /pubmed/30504793 http://dx.doi.org/10.1038/s41598-018-35727-3 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Zuckermann, Marc
Hlevnjak, Mario
Yazdanparast, Haniyeh
Zapatka, Marc
Jones, David T. W.
Lichter, Peter
Gronych, Jan
A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title_full A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title_fullStr A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title_full_unstemmed A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title_short A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
title_sort novel cloning strategy for one-step assembly of multiplex crispr vectors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6269432/
https://www.ncbi.nlm.nih.gov/pubmed/30504793
http://dx.doi.org/10.1038/s41598-018-35727-3
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