Open AccessEfficient CRISPR ‐based genome editing using tandem guide RNA s and editable surrogate reporters
Author(s) -
Liu Wuqing,
Li Shifeng,
Zhang Yunbin,
Li Jinsong,
Li Yiping
Publication year - 2018
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12437
Subject(s) - crispr , genome editing , guide rna , computational biology , rna , tandem , palindrome , gene , biology , genetics , materials science , composite material
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 RNA s 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.