CRISPR directed evolution of the spliceosome for resistance to splicing inhibitors

被引：90
作者：
Butt, Haroon [1 ]
Eid, Ayman [1 ]
Momin, Afaque A. [2 ]
Bazin, Jeremie [3 ]
Crespi, Martin [3 ]
Arold, Stefan T. [2 ]
Mahfouz, Magdy M. [1 ]
机构：
[1] KAUST, Lab Genome Engn & Synthet Biol, Thuwal, Saudi Arabia
[2] KAUST, Div Biol & Environm Sci & Engn, Computat Biosci Res Ctr, Thuwal, Saudi Arabia
[3] Univ Paris Saclay, Univ Paris Diderot, Univ Evry,Sorbonne Paris Cite, Univ Paris Sud,INRA,CNRS,Inst Plant Sci Paris Sac, Orsay, France
来源：
GENOME BIOLOGY | 2019年 / 20卷 / 1期
关键词：
Directed evolution; Genome engineering; Spliceosome; SF3B complex; SF3B1; Splicing modulators; Herboxidiene; Pladienolide B; Spliceostatin A; CRISPR/Cas9; Herbicide resistance; TARGET BASE; RICE; RNA;
D O I：
10.1186/s13059-019-1680-9
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Increasing genetic diversity via directed evolution holds great promise to accelerate trait development and crop improvement. We developed a CRISPR/Cas-based directed evolution platform in plants to evolve the rice (Oryza sativa) SF3B1 spliceosomal protein for resistance to splicing inhibitors. SF3B1 mutant variants, termed SF3B1-GEX1A-Resistant (SGR), confer variable levels of resistance to splicing inhibitors. Studies of the structural basis of the splicing inhibitor binding to SGRs corroborate the resistance phenotype. This directed evolution platform can be used to interrogate and evolve the molecular functions of key biomolecules and to engineer crop traits for improved performance and adaptation under climate change conditions.
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页数：9
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