Manipulating mRNA splicing by base editing in plants

Xue, Chenxiao; Zhang, Huawei; Lin, Qiupeng; Fan, Rong; Gao, Caixia

doi:10.1007/s11427-018-9392-7

Cited by 55 publications

(39 citation statements)

References 23 publications

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“…This study demonstrated that gene knockout can be achieved in New Phytologist using ABE or CBE in Arabidopsis (Kang et al, 2018;Xue et al, 2018). Kang et al (2018) reported that ABE could be harnessed to disrupt the A Á T base pair in an intron acceptor site of Arabidopsis PDS3 gene to cause gene dysfunction.…”

Section: Discussionmentioning

confidence: 87%

“…This study demonstrated that gene knockout can be achieved in both Arabidopsis and rice by mutating the G·C base pair in either the intron donor site GT or acceptor site AG using a CBE independent of DSB generation. Very recently, two other plant groups have demonstrated similar strategies for gene knockout using ABE or CBE in Arabidopsis (Kang et al ., ; Xue et al ., ). Kang et al .…”

Section: Discussionmentioning

confidence: 97%

“…In this study, we deliberately targeted and destroyed the intron splice sites for plant genes without alternative splicing and could observe the induction of new aberrant splicing (shorter transcripts in Figs d, c). In contrast, elimination of the intron splice sites for Arabidopsis genes with alternative splicing could not induce any new aberrant splicing (Xue et al ., ). These complementary findings provide important experimental evidence to support the hypothesis that alternative splicing may be evolved from aberrant splicing when ancient eukaryotic cells are forced to use and benefit from aberrant splicing under certain growth or stressed conditions (Irimia et al ., ).…”

Section: Discussionmentioning

confidence: 97%

“…Notably, PDS3 knockout in that study was also found in a mosaic pattern in transgenic T 1 Arabidopsis plants expressing ABE under the RPS5A promoter. Xue et al (2018) reported that CBE could also be utilized to disrupt an intron donor site to eliminate a specific alternatively spliced gene isoform to study its function in Arabidopsis. Taken together, these studies further expand the applications of base editing technologies in plants by opening up a new possibility for gene knockout.…”

Section: Discussionmentioning

confidence: 99%

“…By mutating the intron splice sites of Arabidopsis genes using ABE or CBE, Arabidopsis null alleles can be obtained in the T 2 generation (Fig. 1) (Kang et al, 2018;Xue et al, 2018). For the first time, we have expanded the applications of such a strategy to rice gene knockout and obtained rice double-null alleles in the T 0 generation (Figs 2, 3).…”

Section: Discussionmentioning

confidence: 99%

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Gene disruption through base editing‐induced messenger RNA missplicing in plants

Xiong

Wang

et al. 2019

New Phytologist

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Summary Gene knockout tools are highly desirable for basic and applied plant research. Here, we leverage the Cas9‐derived cytosine base editor to introduce precise C‐to‐T mutations to disrupt the highly conserved intron donor site GT or acceptor site AG, thereby inducing messenger RNA (mRNA) missplicing and gene disruption. As proof of concept, we successfully obtained Arabidopsis null mutant of MTA gene in the T2 generation and rice double null mutant of GL1‐1 and NAL1 genes in the T0 generation by this strategy. Elimination of the original intron donor site or acceptor site could trigger aberrant splicing at a new specific exonic site, but not at the closest GT or AG site, suggesting cryptic rules governing splice site recognition. The strategy presented expands the applications of base editing technologies in plants by providing a new means for gene inactivation without generating DNA double‐strand breaks, and it can potentially serve as a useful tool for studying the biology of mRNA splicing.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

Gene disruption through base editing‐induced messenger RNA missplicing in plants

Xiong

Wang

et al. 2019

New Phytologist

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CRISPR-Cas Based Precision Breeding in Date Palm: Future Applications

Sattar

Iqbal

Al–Khayri

2021

The Date Palm Genome, Vol. 2

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Base Editing and Prime Editing

Kaya

2023

A Roadmap for Plant Genome Editing

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The development of new adaptations of CRISPR-based genome editing platforms, such as base editing and prime editing, made it possible to broaden the scope and applications of genome editing in plants. First base editing and, more recently, prime editing evade the creation of double-stranded breaks in deoxyribonucleic acid (DNA) and the requirement of donor template of DNA for repair while enhancing editing efficiency and product purity over CRISPR/Cas9. As base-pair changes in genomic DNA determine many significant agronomic traits, crop varieties can be developed by precisely converting specific single bases in plant genomes. While base editing can introduce specific nucleotide changes, such as transition and transversion mutations in the targeted region, prime editing can create precise insertions, deletions, and all 12 types of point mutations using the “search-and-replace” method.This chapter provides the basic principles of base editing and prime editing technologies and their practical applications in plants. The chapter also summarizes the recent breakthroughs in applying base and prime editors in diverse plant species, including their use in improving disease resistance, herbicide resistance, nutritional quality, crop yield, and quality. Finally, this chapter aims to clearly understand base editing and prime editing in plants by outlining potential developments.

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Manipulating mRNA splicing by base editing in plants

Cited by 55 publications

References 23 publications

Gene disruption through base editing‐induced messenger RNA missplicing in plants

Gene disruption through base editing‐induced messenger RNA missplicing in plants

CRISPR-Cas Based Precision Breeding in Date Palm: Future Applications

Base Editing and Prime Editing

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