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


ABSTRACT: 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.

SUBMITTER: Liu W 

PROVIDER: S-EPMC6026697 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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

Liu Wuqing W   Li Shifeng S   Zhang Yunbin Y   Li Jinsong J   Li Yiping Y  

FEBS open bio 20180613 7


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  ...[more]

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