High-efficiency and multiplex adenine base editing in plants using new TadA variants

Yan, Daqi; Ren, Bin; Liü, Lang; Yan, Fang; Li, Shaofang; Wang, Guirong; Sun, Wenxian; Zhou, Xueping; Zhou, Huanbin

doi:10.1016/j.molp.2021.02.007

Cited by 83 publications

(87 citation statements)

References 32 publications

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“…We further used three sgRNA expression cassettes to investigate the effects of sgRNA expression cassette on the editing efficiency of ABE8e ( Figure 3 a). The expression cassette of ABE8e is a regular expression cassette used in previous studies [ 14 , 15 , 19 ]. The cassette of STU-ABE8e combined the sgRNA and Cas9 into a single transcript unit (STU) ( Figure 3 a).…”

Section: Resultsmentioning

confidence: 99%

“…The efficiency of ABE8e is significantly improved in human cells, especially at poorly edited targets with ABE7.10, which could provide a useful tool for broadening adenine base editing capability [ 12 ]. While we prepared this manuscript, the ABE8e has been reported in rice [ 14 , 15 , 16 ]. However, the comparison between ABE7.10 and ABE8e and the effects of the sgRNA expression system on ABE8e have not been investigated.…”

Section: Introductionmentioning

confidence: 99%

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ABE8e with Polycistronic tRNA-gRNA Expression Cassette Sig-Nificantly Improves Adenine Base Editing Efficiency in Nicotiana benthamiana

Wang

Xie

et al. 2021

IJMS

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Adenine base editor containing TadA8e (ABE8e) has been reported in rice. However, the application of ABE8e in other plant species has not been described, and the comparison between ABE8e and ABE7.10, which is widely used in plants, has also been poorly studied. Here, we developed the ABE8e with the polycistronic tRNA-gRNA expression cassette (PTG-ABE8e) and PTG-ABE7.10 and compared their A-to-G editing efficiencies using both transient and stable transformation in the allotetraploid Nicotiana benthamiana. We found that the editing efficiency of PTG-ABE8e was significantly higher than that of PTG-ABE7.10, indicating that ABE8e was more efficient for A-to-G conversion in N. benthamiana. We further optimized the ABE8e editing efficiency by changing the sgRNA expression cassette and demonstrated that both PTG and single transcript unit (STU) enhanced ABE8e efficiency for A-to-G conversion in N. benthamiana. We also estimated the potential off-target effect of PTG-ABE8e at potential off-targeting sites predicted using an online tool in transgenic plants, and no off-target editing event was found for potential off-targeting sites selected, indicating that ABE8e could specifically facilitate A-to-G conversion. Our results showed that ABE8e with PTG structure was more suitable for A-to-G conversion in N. benthamiana and provided valuable clues for optimizing ABE tools in other plants.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

ABE8e with Polycistronic tRNA-gRNA Expression Cassette Sig-Nificantly Improves Adenine Base Editing Efficiency in Nicotiana benthamiana

Wang

Xie

et al. 2021

IJMS

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“…These exhibited increased editing efficiencies in human cells. Out of these editors, TadA8e has the highest efficiency of creating A-to-G changes across different target sites ( Yan et al, 2021 ). Yan et al (2021) developed an improved base editor, named as TadA9, by introducing V82S/Q154R mutations in TadA8e.…”

Section: Currently Available Types Of Base Editorsmentioning

confidence: 99%

“…Out of these editors, TadA8e has the highest efficiency of creating A-to-G changes across different target sites ( Yan et al, 2021 ). Yan et al (2021) developed an improved base editor, named as TadA9, by introducing V82S/Q154R mutations in TadA8e. TadA9 was determined to be very compatible with various nickase systems, such as CRISPR/SpCas9, CRISPR/SpCas9-NG, CRISPR/SpRY, and CRISPR/ScCas9, and it exhibited high efficiency in editing four herbicide target genes in commercial rice.…”

Section: Currently Available Types Of Base Editorsmentioning

confidence: 99%

Base Editing in Plants: Applications, Challenges, and Future Prospects

Azameti

Palnam

2021

Front. Plant Sci.

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The ability to create targeted modifications in the genomes of plants using genome editing technologies has revolutionized research in crop improvement in the current dispensation of molecular biology. This technology has attracted global attention and has been employed in functional analysis studies in crop plants. Since many important agronomic traits are confirmed to be determined by single-nucleotide polymorphisms, improved crop varieties could be developed by the programmed and precise conversion of targeted single bases in the genomes of plants. One novel genome editing approach which serves for this purpose is base editing. Base editing directly makes targeted and irreversible base conversion without creating double-strand breaks (DSBs). This technology has recently gained quick acceptance and adaptation because of its precision, simplicity, and multiplex capabilities. This review focuses on generating different base-editing technologies and how efficient they are in editing nucleic acids. Emphasis is placed on the exploration and applications of these base-editing technologies to enhance crop production. The review also highlights the drawbacks and the prospects of this new technology.

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“…Additionally, the mutation of the OsTubA2 gene can also be rapidly introduced to confer resistance to both trifluralin and pendimethalin herbicides in rice using CRIPSR-mediated adenine base editors [73]. Recently, Yan et al [79] simultaneously developed novel SNPs in four endogenous herbicide target genes (OsALS1, OsGS1, OsTubA2, OsACC) with induced efficient A-to-G conversion using new TadA variants, named TadA9.…”

Section: Improving Herbicide Resistance Through Base Editingmentioning

confidence: 99%

The Development of Herbicide Resistance Crop Plants Using CRISPR/Cas9-Mediated Gene Editing

Dong

Huang

Wang

2021

Genes

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The rapid increase in herbicide-resistant weeds creates a huge challenge to global food security because it can reduce crop production, causing considerable losses. Combined with a lack of novel herbicides, cultivating herbicide-resistant crops becomes an effective strategy to control weeds because of reduced crop phytotoxicity, and it expands the herbicidal spectrum. Recently developed clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas)-mediated genome editing techniques enable efficiently targeted modification and hold great potential in creating desired plants with herbicide resistance. In the present review, we briefly summarize the mechanism responsible for herbicide resistance in plants and then discuss the applications of traditional mutagenesis and transgenic breeding in cultivating herbicide-resistant crops. We mainly emphasize the development and use of CRISPR/Cas technology in herbicide-resistant crop improvement. Finally, we discuss the future applications of the CRISPR/Cas system for developing herbicide-resistant crops.

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High-efficiency and multiplex adenine base editing in plants using new TadA variants

Cited by 83 publications

References 32 publications

ABE8e with Polycistronic tRNA-gRNA Expression Cassette Sig-Nificantly Improves Adenine Base Editing Efficiency in Nicotiana benthamiana

ABE8e with Polycistronic tRNA-gRNA Expression Cassette Sig-Nificantly Improves Adenine Base Editing Efficiency in Nicotiana benthamiana

Base Editing in Plants: Applications, Challenges, and Future Prospects

The Development of Herbicide Resistance Crop Plants Using CRISPR/Cas9-Mediated Gene Editing

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