PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies

Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple...

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Detalles Bibliográficos
Autores principales: Yuan, Guoliang, Martin, Stanton, Hassan, Md Mahmudul, Tuskan, Gerald A., Yang, Xiaohan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Communication
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406951/
https://www.ncbi.nlm.nih.gov/pubmed/36010544
http://dx.doi.org/10.3390/cells11162467
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author Yuan, Guoliang
Martin, Stanton
Hassan, Md Mahmudul
Tuskan, Gerald A.
Yang, Xiaohan
author_facet Yuan, Guoliang
Martin, Stanton
Hassan, Md Mahmudul
Tuskan, Gerald A.
Yang, Xiaohan
author_sort Yuan, Guoliang
collection PubMed
description Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple guide RNAs (gRNAs) into a CRISPR vector with up to 18 gRNAs. Here, we demonstrate that PARA is capable of the efficient assembly of transfer RNA/Csy4/ribozyme-based gRNA arrays. To aid in this process and to streamline vector construction, we developed a user-friendly PARAweb tool for designing PCR primers and component DNA parts and simulating assembled gRNA arrays and vector sequences.
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spelling pubmed-94069512022-08-26 PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies Yuan, Guoliang Martin, Stanton Hassan, Md Mahmudul Tuskan, Gerald A. Yang, Xiaohan Cells Communication Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple guide RNAs (gRNAs) into a CRISPR vector with up to 18 gRNAs. Here, we demonstrate that PARA is capable of the efficient assembly of transfer RNA/Csy4/ribozyme-based gRNA arrays. To aid in this process and to streamline vector construction, we developed a user-friendly PARAweb tool for designing PCR primers and component DNA parts and simulating assembled gRNA arrays and vector sequences. MDPI 2022-08-09 /pmc/articles/PMC9406951/ /pubmed/36010544 http://dx.doi.org/10.3390/cells11162467 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Yuan, Guoliang
Martin, Stanton
Hassan, Md Mahmudul
Tuskan, Gerald A.
Yang, Xiaohan
PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title_full PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title_fullStr PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title_full_unstemmed PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title_short PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
title_sort para: a new platform for the rapid assembly of grna arrays for multiplexed crispr technologies
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406951/
https://www.ncbi.nlm.nih.gov/pubmed/36010544
http://dx.doi.org/10.3390/cells11162467
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