Harnessing the native type I-B CRISPR-Cas for genome editing in a polyploid archaeon

Feiyue Cheng^{a, b},
Luyao Gong^{a, b},
Dahe Zhao^{a, b},
Haibo Yang^{a, b},
Jian Zhou^a,
Ming Li^{a
, ,},
Hua Xiang^{a, b
, ,}

a.
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
b.
University of Chinese Academy of Sciences, Beijing 100049, China

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Corresponding author: E-mail address: lim_im@im.ac.cn (Ming Li); E-mail address: xiangh@im.ac.cn (Hua Xiang)

摘要

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Abstract: Research on CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR associated protein) systems has led to the revolutionary CRISPR/Cas9 genome editing technique. However, for most archaea and half of bacteria, exploitation of their native CRISPR-Cas machineries may be more straightforward and convenient. In this study, we harnessed the native type I-B CRISPR-Cas system for precise genome editing in the polyploid haloarchaeon Haloarcula hispanica. After testing different designs, the editing tool was optimized to be a single plasmid that carries both the self-targeting mini-CRISPR and a 600–800 bp donor. Significantly, chromosomal modifications, such as gene deletion, gene tagging or single nucleotide substitution, were precisely introduced into the vast majority of the transformants. Moreover, we showed that simultaneous editing of two genomic loci could also be readily achieved by one step. In summary, our data demonstrate that the haloarchaeal CRISPR-Cas system can be harnessed for genome editing in this polyploid archaeon, and highlight the convenience and efficiency of the native CRISPR-based genome editing strategy.
- CRISPR-Cas /
- Genome editing /
- Polyploid

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doi: 10.1016/j.jgg.2017.09.010

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Harnessing the native type I-B CRISPR-Cas for genome editing in a polyploid archaeon

Corresponding author: E-mail address: lim_im@im.ac.cn (Ming Li); E-mail address: xiangh@im.ac.cn (Hua Xiang)

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doi: 10.1016/j.jgg.2017.09.010

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Harnessing the native type I-B CRISPR-Cas for genome editing in a polyploid archaeon

Corresponding author: E-mail address: lim_im@im.ac.cn (Ming Li); E-mail address: xiangh@im.ac.cn (Hua Xiang)

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