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Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics

The CRISPR diagnostic (CRISPR-Dx) technology that employs the trans-cleavage activities has shown great potential in diagnostic sensitivity, specificity, convenience, and portability, and has been recognized as the next-generation diagnostic methods. However, due to the lack of standardized definiti...

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Autores principales: Lv, Hailong, Wang, Jian, Zhang, Jian, Chen, Yijian, Yin, Lei, Jin, Dian, Gu, Dayong, Zhao, Huailong, Xu, Yong, Wang, Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8667580/
https://www.ncbi.nlm.nih.gov/pubmed/34912315
http://dx.doi.org/10.3389/fmicb.2021.766464
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author Lv, Hailong
Wang, Jian
Zhang, Jian
Chen, Yijian
Yin, Lei
Jin, Dian
Gu, Dayong
Zhao, Huailong
Xu, Yong
Wang, Jin
author_facet Lv, Hailong
Wang, Jian
Zhang, Jian
Chen, Yijian
Yin, Lei
Jin, Dian
Gu, Dayong
Zhao, Huailong
Xu, Yong
Wang, Jin
author_sort Lv, Hailong
collection PubMed
description The CRISPR diagnostic (CRISPR-Dx) technology that employs the trans-cleavage activities has shown great potential in diagnostic sensitivity, specificity, convenience, and portability, and has been recognized as the next-generation diagnostic methods. However, due to the lack of standardized definition of Cas trans-cleavage enzymatic units, it is difficult to standardize the present CRISPR-Dx systems, which have undoubtedly impeded the development of the CRISPR-Dx industry. To solve the problem, we here first systematically optimized the reaction systems for Cas12a, and then defined its trans-cleavage units (transU), which we believe will be of great importance and interest to researchers in both molecular diagnostic industry and basic research. Moreover, a simple protocol was provided to facilitate a step-by-step measurement of the Cas12a transU, which can also act as a reference for the definition of the transU for other Cas proteins.
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spelling pubmed-86675802021-12-14 Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics Lv, Hailong Wang, Jian Zhang, Jian Chen, Yijian Yin, Lei Jin, Dian Gu, Dayong Zhao, Huailong Xu, Yong Wang, Jin Front Microbiol Microbiology The CRISPR diagnostic (CRISPR-Dx) technology that employs the trans-cleavage activities has shown great potential in diagnostic sensitivity, specificity, convenience, and portability, and has been recognized as the next-generation diagnostic methods. However, due to the lack of standardized definition of Cas trans-cleavage enzymatic units, it is difficult to standardize the present CRISPR-Dx systems, which have undoubtedly impeded the development of the CRISPR-Dx industry. To solve the problem, we here first systematically optimized the reaction systems for Cas12a, and then defined its trans-cleavage units (transU), which we believe will be of great importance and interest to researchers in both molecular diagnostic industry and basic research. Moreover, a simple protocol was provided to facilitate a step-by-step measurement of the Cas12a transU, which can also act as a reference for the definition of the transU for other Cas proteins. Frontiers Media S.A. 2021-11-29 /pmc/articles/PMC8667580/ /pubmed/34912315 http://dx.doi.org/10.3389/fmicb.2021.766464 Text en Copyright © 2021 Lv, Wang, Zhang, Chen, Yin, Jin, Gu, Zhao, Xu and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Lv, Hailong
Wang, Jian
Zhang, Jian
Chen, Yijian
Yin, Lei
Jin, Dian
Gu, Dayong
Zhao, Huailong
Xu, Yong
Wang, Jin
Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title_full Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title_fullStr Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title_full_unstemmed Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title_short Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics
title_sort definition of crispr cas12a trans-cleavage units to facilitate crispr diagnostics
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8667580/
https://www.ncbi.nlm.nih.gov/pubmed/34912315
http://dx.doi.org/10.3389/fmicb.2021.766464
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