Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters

Cleavage efficiency plays a key role in clustered regularly interspaced short palindromic repeat (CRISPR)‐based gene editing, particularly when the given guide RNA exhibits low cleavage activity. Here, we describe the packaging of tandem guide RNAs and single‐strand annealing‐based surrogate reporte...

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Detalles Bibliográficos
Autores principales: Liu, Wuqing, Li, Shifeng, Zhang, Yunbin, Li, Jinsong, Li, Yiping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Methods
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6026697/
https://www.ncbi.nlm.nih.gov/pubmed/29988563
http://dx.doi.org/10.1002/2211-5463.12437
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author Liu, Wuqing
Li, Shifeng
Zhang, Yunbin
Li, Jinsong
Li, Yiping
author_facet Liu, Wuqing
Li, Shifeng
Zhang, Yunbin
Li, Jinsong
Li, Yiping
author_sort Liu, Wuqing
collection PubMed
description Cleavage efficiency plays a key role in clustered regularly interspaced short palindromic repeat (CRISPR)‐based gene editing, particularly when the given guide RNA exhibits low cleavage activity. Here, we describe the packaging of tandem guide RNAs and single‐strand annealing‐based surrogate reporter cassettes into the CRISPR/CRISPR‐associated protein 9 vector, which increased gene‐editing efficiency by 4.94–6.31‐fold and simultaneously enriched the proportion of genetically modified cells. This strategy may substantially improve genome‐editing efficiency for demanding applications.
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spelling pubmed-60266972018-07-09 Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters Liu, Wuqing Li, Shifeng Zhang, Yunbin Li, Jinsong Li, Yiping FEBS Open Bio Methods Cleavage efficiency plays a key role in clustered regularly interspaced short palindromic repeat (CRISPR)‐based gene editing, particularly when the given guide RNA exhibits low cleavage activity. Here, we describe the packaging of tandem guide RNAs and single‐strand annealing‐based surrogate reporter cassettes into the CRISPR/CRISPR‐associated protein 9 vector, which increased gene‐editing efficiency by 4.94–6.31‐fold and simultaneously enriched the proportion of genetically modified cells. This strategy may substantially improve genome‐editing efficiency for demanding applications. John Wiley and Sons Inc. 2018-06-13 /pmc/articles/PMC6026697/ /pubmed/29988563 http://dx.doi.org/10.1002/2211-5463.12437 Text en © 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods
Liu, Wuqing
Li, Shifeng
Zhang, Yunbin
Li, Jinsong
Li, Yiping
Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title_full Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title_fullStr Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title_full_unstemmed Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title_short Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters
title_sort efficient crispr‐based genome editing using tandem guide rnas and editable surrogate reporters
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6026697/
https://www.ncbi.nlm.nih.gov/pubmed/29988563
http://dx.doi.org/10.1002/2211-5463.12437
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AT lijinsong efficientcrisprbasedgenomeeditingusingtandemguidernasandeditablesurrogatereporters
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